Macrocyclic compounds as trk kinase inhibitors

ABSTRACT

Compounds of Formula I: and pharmaceutically acceptable salts thereof, wherein ring A, ring B, W, m, D, R 2 , R 2a , R 3 , R 3a , and Z are as defined herein, are inhibitors of Trk kinases and are useful in the treatment of pain, cancer, inflammation, neurodegenerative diseases and certain infectious diseases.

The present invention relates to novel compounds, to pharmaceuticalcompositions comprising the compounds, to processes for making thecompounds and to the use of the compounds in therapy. More particularly,it relates to certain macrocyclic compounds which exhibit Trk familyprotein tyrosine kinase inhibition, and which are useful in thetreatment of pain, cancer, inflammation, neurodegenerative diseases andcertain infectious diseases.

The current treatment regimes for pain conditions utilize severalclasses of compounds. The opioids (such as morphine) have severaldrawbacks including emetic, constipatory and negative respiratoryeffects, as well as the potential for addiction. Non-steroidalanti-inflammatory analgesics (NSAIDs, such as COX-1 or COX-2 types) alsohave drawbacks including insufficient efficacy in treating severe painand the potential for internal gastrointestinal bleeding. In addition,COX-1 inhibitors can cause ulcers of the mucosa. Accordingly, there is acontinuing need for new and more effective treatments for the relief ofpain, especially chronic pain.

Trk's are high affinity receptor tyrosine kinases activated by a groupof soluble growth factors called neurotrophins (NT). The Trk receptorfamily has three members: TrkA, TrkB and TrkC. Among the neurotrophinsare (i) nerve growth factor (NGF) which activates TrkA, (ii)brain-derived neurotrophic factor (BDNF) and NT-4/5 which activate TrkBand (iii) NT3 which activates TrkC. Trk's are widely expressed inneuronal tissue and are implicated in the maintenance, signaling andsurvival of neuronal cells (Patapoutian, A. et al., Current Opinion inNeurobiology, 2001, 11, 272-280).

Inhibitors of the Trk/neurotrophin pathway have been demonstrated to beeffective in numerous pre-clinical animal models of pain. For example,antagonistic NGF and TrkA antibodies such as RN-624 have been shown tobe efficacious in inflammatory and neuropathic pain animal models(Woolf, C. J. et al. (1994) Neuroscience 62,327-331; Zahn, P. K. et al.(2004) J. Pain 5, 157-163; McMahon, S. B. et al., (1995) Nat. Med. 1,774-780; Ma, Q. P. and Woolf, C. J. (1997) Neuroreport 8, 807-810;Shelton, D. L. et al. (2005) Pain 116, 8-16; Delafoy, L. et al. (2003)Pain 105, 489-497; Lamb, K. et al. (2003) Neurogastroenterol. Motil. 15,355-361; Jaggar, S. I. et al. (1999) Br. J. Anaesth. 83, 442-448) andneuropathic pain animal models (Ramer, M. S. and Bisby, M. A. (1999)Eur. J. Neurosci. 11, 837-846; Ro, L. S. et al. (1999); Pain 79, 265-274Herzberg, U. et al. (1997) Neuroreport 8, 1613-1618; Theodosiou, M. etal. (1999) Pain 81, 245-255; Li, L. et al. (2003) Mol. Cell. Neurosci.23, 232-250; Gwak, Y. S. et al. (2003) Neurosci. Lett. 336, 117-120).

It has also been shown that NGF secreted by tumor cells and tumorinvading macrophages directly stimulates TrkA located on peripheral painfibers. Using various tumor models in both mice and rats, it wasdemonstrated that neutralizing NGF with a monoclonal antibody inhibitscancer related pain to a degree similar or superior to the highesttolerated dose of morphine. In addition, activation of the BDNF/TrkBpathway has been implicated in numerous studies as a modulator ofvarious types of pain including inflammatory pain (Matayoshi, S., J.Physiol. 2005, 569:685-95), neuropathic pain (Thompson, S. W., Proc.Natl. Acad. Sci. USA 1999, 96:7714-18) and surgical pain (Li, C.-Q. etal., Molecular Pain, 2008, 4(28), 1-11).

Recent literature has also shown that overexpression, activation,amplification and/or mutation of Trk kinases are associated with manycancers including neuroblastoma (Brodeur, G. M., Nat. Rev. Cancer 2003,3, 203-216), ovarian (Davidson. B., et al., Clin. Cancer Res. 2003, 9,2248-2259), colorectal cancer (Bardelli, A., Science 2003, 300, 949),melanoma (Truzzi, F., et al., Dermato-Endocrinology 2008, 3 (1), pp.32-36), head and neck cancer (Yilmaz, T., et al., Cancer Biology andTherapy 2010, 10 (6), pp. 644-653), gastric carcinoma (Du, J. et al.,World Journal of Gastroenterology 2003, 9 (7), pp. 1431-1434), lungcarcinoma (Ricci A., et al., American Journal of Respiratory Cell andMolecular Biology 25 (4), pp. 439-446), breast cancer (Jin, W., et al.,Carcinogenesis 2010, 31 (11), pp. 1939-1947), Glioblastoma (Wadhwa, S.,et al., Journal of Biosciences 2003, 28 (2), pp. 181-188),medulloblastoma (Gruber-Olipitz, M., et al., Journal of ProteomeResearch 2008, 7 (5), pp. 1932-1944), secratory breast cancer (Euthus,D. M., et al., Cancer Cell 2002, 2 (5), pp. 347-348), salivary glandcancer (Li, Y.-G., et al., Chinese Journal of Cancer Prevention andTreatment 2009, 16 (6), pp. 428-430), papillary thyroid carcinoma(Greco, A., et al., Molecular and Cellular Endocrinology 2010, 321 (1),pp. 44-49) and adult myeloid leukemia (Eguchi, M., et al., Blood 1999,93 (4), pp. 1355-1363). In preclinical models of cancer, non-selectivesmall molecule inhibitors of Trk A, B and C were efficacious in bothinhibiting tumor growth and stopping tumor metastasis (Nakagawara, A.(2001) Cancer Letters 169:107-114; Meyer, J. et al. (2007) Leukemia,1-10; Pierottia, M. A. and Greco A., (2006) Cancer Letters 232:90-98;Eric Adriaenssens, E., et al. Cancer Res (2008) 68:(2) 346-351).

In addition, inhibition of the neurotrophin/Trk pathway has been shownto be effective in treatment of pre-clinical models of inflammatorydiseases with NGF antibodies or non-selective small molecule inhibitorsof Trk A, B and C. For example, inhibition of the neurotrophin/Trkpathway has been implicated in preclinical models of inflammatory lungdiseases including asthma (Freund-Michel, V; Frossard, N., Pharmacology& Therapeutics (2008), 117(1), 52-76), interstitial cystitis (Hu Vivian,Y., et. al. The Journal of Urology (2005), 173(3), 1016-21),inflammatory bowel diseases including ulcerative colitis and Crohn'sdisease (Di Mola, F. F., et. al., Gut (2000), 46(5), 670-678) andinflammatory skin diseases such as atopic dermatitis (Dou, Y.-C., et.al. Archives of Dermatological Research (2006), 298(1), 31-37), eczemaand psoriasis (Raychaudhuri, S. P., et al., J. Investigative Dermatology(2004), 122(3), 812-819).

The neurotrophirv/Trk pathway, particularly BDNF/TrkB, has also beenimplicated in the etiology of neurodegenerative diseases includingmultiple sclerosis, Parkinson's disease and Alzheimer's Disease(Sohrabji, F., Lewis, Danielle K., Frontiers in Neuroendocrinology(2006), 27(4), 404-414).

The TrkA receptor is also thought to be critical to the disease processin the infection of the parasitic infection of Trypanosoma cruzi (Chagasdisease) in human hosts (de Melo-Jorge, M., et al., Cell Host & Microbe(2007), 1(4), 251-261).

Several classes of small molecule inhibitors of Trk kinases said to beuseful for treating pain or cancer are known (Expert Opin. Ther. Patents(2009) 19(3)).

There remains a need, however, for compounds and methods for thetreatment of pain, in particular chronic pain, as well as for thetreatment of cancer, inflammation, neurodegenerative diseases andcertain infectious diseases.

SUMMARY OF THE INVENTION

It has now been found that macrocyclic compounds are inhibitors of Trkkinases, in particular inhibitors of TrkA and/or TrkB and/or TrkC, andare useful for treating disorders and diseases such as cancer and pain,including chronic and acute pain. Compounds which are inhibitors of TrkAand/or TrkB may be useful in the treatment of multiple types of painincluding inflammatory pain, neuropathic pain, and pain associated withcancer, surgery and bone fracture. In addition, compounds of theinvention may be useful for treating inflammation, neurodegenerativediseases and certain infectious diseases.

Accordingly, in one aspect present invention provides novel compoundshaving the general Formula I:

and stereoisomers and pharmaceutically acceptable salts and solvatesthereof, wherein ring A, ring B, W, m, D, R², R^(2a), R³, R^(3a), and Zare as defined herein.

In another aspect, the present invention provides novel compounds havingthe general Formula I:

or pharmaceutically acceptable salts or solvates thereof, wherein ringA, W, m, R², R^(2a), R³, Z, R⁵ and R⁶ are as defined herein.

In another aspect of the invention, there are provided pharmaceuticalcompositions comprising compounds of Formula I and a carrier, diluent orexcipient.

In another aspect of the invention, there is provided a method fortreating or preventing pain, cancer, inflammation, neurodegenerativediseases and certain infectious diseases in a mammal, comprisingadministering to said mammal an effective amount of a compound ofFormula I.

In another aspect of the invention, there is provided a use of acompound of Formula I in the manufacture of a medicament for thetreatment or prevention of pain, cancer, inflammation, neurodegenerativediseases and certain infectious diseases.

In another aspect of the invention, there is provided a use of acompound of Formula I in the treatment or prevention of pain, cancer,inflammation, neurodegenerative diseases and certain infectiousdiseases.

Another aspect provides intermediates for preparing compounds of FormulaI. In one embodiment, certain compounds of Formula I may be used asintermediates for the preparation of other compounds of Formula I.

Another aspect includes processes for preparing, methods of separation,and methods of purification of the compounds described herein.

DETAILED DESCRIPTION OF THE INVENTION

One embodiment of this invention provides compounds of the generalFormula I containing a pyrazolo[1,5-a]pyrimidinyl ring and having thestructure:

or pharmaceutically acceptable salts or solvates thereof, wherein:

ring A is selected from rings A-1, A-2 and A-3 having the structures:

wherein the wavy line labeled 1 indicates the point of attachment ofring A to ring B and the wavy line labeled 2 indicates the point ofattachment of ring A to W;

X is N or CH;

Y is H or F;

R¹ is H, (1-3C)alkoxy or halogen;

ring B is selected from rings B-1 and B-2 having the structures:

wherein the wavy line labeled 3 indicates the point of attachment toring A and the wavy line labeled 4 indicates the point of attachment tothe pyrazolo[1,5-a]pyrimidine ring of Formula I;

W is O, NH or CH₂, wherein when ring A is A-2, then W is CH₂;

m is 0, 1 or 2;

D is carbon;

R² and R^(2a) are independently H, F, (1-3C)alkyl or OH, provided thatR² and R^(2a) are not both OH;

R³ and R^(3a) are independently H, (1-3C)alkyl or hydroxy(1-3C)alkyl;

or D is carbon or nitrogen, R² and R³ are absent and R^(2a) and R^(3a)together with the atoms to which they are attached form a 5-6 memberedheteroaryl ring having 1-2 ring heteroatoms;

Z is *—NR^(4a)C(═O)—, *—ONHC(═O)—, *—NR^(4b)CH₂— or *—OC(═O)—, whereinthe asterisk indicates the point of attachment of Z to the carbonbearing R³;

R^(4a) is H, (1-6C)alkyl, fluoro(1-6C)alkyl, difluoro(1-6C)alkyl,trifluoro(1-6C)alkyl, hydroxy(1-6C alkyl) or dihydroxy(2-6C alkyl);

R^(4b) is H, (1-6C)alkyl, fluoro(1-6C)alkyl, difluoro(1-6C)alkyl,trifluoro(1-6C)alkyl, hydroxy(1-6C alkyl), dihydroxy(2-6C alkyl), (1-6Calkyl)C(O)—, (3-6C cycloalkyl)C(O)—, Ar¹C(O)—, HOCH₂C(O)—, (1-6Calkyl)sulfonyl, (3-6C cycloalkyl)sulfonyl, Ar²(SO₂)—, HO₂CCH₂— or (1-6Calkyl)NH(CO)—;

Ar¹ is phenyl optionally substituted with one or more substituentsindependently selected from halogen, (1-6C)alkyl, and (1-6C)alkoxy;

Ar² is phenyl optionally substituted with one or more substituentsindependently selected from halogen, (1-6C)alkyl, and (1-6C)alkoxy; and

R⁵ and R⁶ are independently H, halogen, OH, (1-6C)alkyl orhydroxy(1-6C)alkyl.

In one embodiment of Formula I, ring B is ring B-2 having the structure:

D is carbon, R² and R^(2a) are independently (1-3C)alkyl, and R³ andR^(3a) are independently H, (1-3C)alkyl or hydroxy(1-3C)alkyl, or

D is carbon or nitrogen, R² and R³ are absent and R^(2a) and R^(3a)together with the atoms to which they are attached form a 5-6 memberedheteroaryl ring having 1-2 ring heteroatoms.

In one embodiment of Formula I, ring A is ring A-1 having the structure

wherein X, Y and R¹ are as defined for Formula I. In one embodiment ofFormula I, X is CH. In one embodiment, X is N. In one embodiment ofFormula I, Y is F. In one embodiment, Y is H. In one embodiment ofFormula I, R¹ is H. In one embodiment, R¹ is (1-3C)alkoxy. A particularexample is methoxy. In one embodiment, R¹ is halogen. In one embodiment,R¹ is F.

Particular examples of ring A when represented by structure A-1 includethe structures:

In one embodiment, ring A is ring A-2 having the structure

wherein Y is H or F. In one embodiment, Y is F. In one embodiment, Y isH. In one embodiment, R¹ is H. In one embodiment, R¹ is (1-3C)alkoxy. Aparticular example is methoxy. In one embodiment, R¹ is halogen. In oneembodiment, R¹ is F.

Particular examples of ring A when represented by ring A-2 are thestructures:

In one embodiment of Formula I, ring A is ring A-3 having the structure

wherein Y and R¹ is as defined for Formula I. In one embodiment, Y is F.In one embodiment, Y is H. In one embodiment, R¹ is H. In oneembodiment, R¹ is (1-3C)alkoxy. A particular example is methoxy. In oneembodiment, R¹ is halogen. In one embodiment, R¹ is F.

Particular examples of ring A when represented by ring A-3 are thestructures:

In one embodiment of Formula I, W is O.

In one embodiment, W is NH.

In one embodiment, W is CH₂.

In one embodiment of Formula I, D is carbon, R² and R^(2a) areindependently H, F, (1-3C)alkyl or OH (provided that R² and R^(2a) arenot both OH), and R³ and R^(3a) are independently H, (1-3C)alkyl orhydroxy(1-3C)alkyl.

In one embodiment, R² and R^(2a) are independently H, F, methyl or OH,provided that R² and R^(2a) are not both OH.

In one embodiment, R² and R^(2a) are both H.

In one embodiment, R² is H and R^(2a) is F.

In one embodiment, R² and R^(2a) are both F.

In one embodiment, R² is H and R^(2a) is OH.

In one embodiment, R² is H and R^(2a) is methyl.

In one embodiment, R² and R^(2a) are both methyl.

In one embodiment, R³ and R^(3a) are independently H, (1-3C)alkyl orhydroxy(1-3C)alkyl.

In one embodiment, R^(3a) is H. In one embodiment, R³ is H. In oneembodiment, both R³ and R^(3a) are H.

In one embodiment, R^(3a) is (1-3C)alkyl. Examples include methyl,ethyl, propyl and isopropyl. In one embodiment, R³ is (1-3C)alkyl.Examples include methyl, ethyl, propyl and isopropyl.

In one embodiment, R^(3a) is (1-3C)alkyl and R³ is H. In one embodiment,R^(3a) is methyl and R³ is H.

In one embodiment, both R^(3a) and R³ are (1-3C)alkyl. In oneembodiment, R^(3a) and R^(3a) are both methyl.

In one embodiment, R³ is hydroxy(1-3C)alkyl. Examples includehydroxymethyl, 2-hydroxyethyl, 2-hydroxypropyl, and 3-hydroxypropyl. Inone embodiment, R³ is hydroxymethyl, 2-hydroxyethyl, 2-hydroxypropyl, or3-hydroxypropyl and R^(3a) is H.

In one embodiment of Formula I, D is carbon or nitrogen, R² and R³ areabsent, and R^(2a) and R^(3a) together with the atoms to which they areattached form a 5-6 membered heteroaryl ring having 1-2 ringheteroatoms. In one embodiment, R^(2a) and R^(3a) together with theatoms to which they are attached form a 5-6 membered heteroaryl ringhaving 1-2 ring nitrogen atoms. Examples of heteroaryl rings includepyridyl and pyrazolyl rings. Specific examples of heteroaryl ringsinclude the structures:

In one embodiment, Z is *—NR^(4a)C(═O)—.

In one embodiment, R^(4a) is H.

In one embodiment, R^(4a) is (1-6C)alkyl. Examples include methyl,ethyl, propyl, isopropyl, butyl, and isobutyl.

In one embodiment, R^(4a) is fluoro(1-6C)alkyl. Examples includefluoromethyl and 2-fluoroethyl.

In one embodiment, R^(4a) is difluoro(1-6C)alkyl. Example includedifluoromethyl and 2,2-difluoroethyl.

In one embodiment, R^(4a) is trifluoro(1-6C)alkyl. Examples includetrifluoromethyl and 2,2,2-trifluoroethyl.

In one embodiment, R^(4a) is hydroxy(1-6C alkyl). Examples includehydroxymethyl, 2-hydroxyethyl, 2-hydroxypropyl and 3-hydroxypropyl.

In one embodiment, R^(4a) is dihydroxy(2-6C alkyl). An example includes2,3-dihydroxypropyl.

In one embodiments, R^(4a) is H or (1-6C)alkyl. In one embodiment,R^(4a) is H or Me.

An example of Z when represented by *—NR^(4a)C(═O)— is *—ONHC(═O)—.

In one embodiment, Z is *—NR^(4b)CH₂—.

In one embodiment, R^(4b) is H.

In one embodiment, R^(4b) is selected from (1-6C)alkyl,fluoro(1-6C)alkyl, difluoro(1-6C)alkyl, and trifluoro(1-6C)alkyl.

In one embodiment, R^(4b) is (1-6C)alkyl. Examples include methyl,ethyl, propyl, isopropyl, butyl and tert-butyl. In one embodiment,R^(4b) is methyl.

In one embodiment, R^(4b) is fluoro(1-6C)alkyl. Examples includefluoromethyl and 2-fluoroethyl.

In one embodiment, R^(4b) is difluoro(1-6C)alkyl. Example includedifluoromethyl and 2,2-difluoroethyl.

In one embodiment, R^(4b) is trifluoro(1-6C)alkyl. Examples includetrifluoromethyl and 2,2,2-trifluoroethyl.

In one embodiment, R^(4b) is selected from (1-6C alkyl)C(O)—, (3-6Ccycloalkyl)C(O)—, Ar¹C(O)— and HOCH₂C(O)—.

In one embodiment, R^(4b) is (1-6C alkyl)C(O)—. Examples includeCH₃C(O)—, CH₃CH₂C(O)—, CH₃CH₂CH₂C(O)—, and (CH₃)₂CHC(O)—. In oneembodiment, R⁴ is CH₃C(O)—.

In one embodiment, R^(4b) is (3-6C cycloalkyl)C(O)—. Examples includecyclopropylC(O)—, cyclobutylC(O)—, cyclopentylC(O)— and cyclohexylC(O)—.

In one embodiment, R^(4b) is Ar¹C(O)—. An example is phenylC(O)—.

In one embodiment, R^(4b) is HOCH₂C(O)—.

In one embodiment, R^(4b) is selected from (1-6C alkyl)sulfonyl, (3-6Ccycloalkyl)sulfonyl, and Ar²(SO₂)—.

In one embodiment, R^(4b) is (1-6C alkyl)sulfonyl. Examples includemethylsulfonyl, ethylsulfonyl and propylsulfonyl.

In one embodiment, R^(4b) is (3-6C cycloalkyl)sulfonyl. Examples includecyclopropylsulfonyl, cyclobutylsulfonyl, cyclopentylsulfonyl andcyclohexylsulfonyl. In one embodiment, R⁴ is methylsulfonyl.

In one embodiment, R^(4b) is Ar²(SO₂)—. An example is phenylsulfonyl.

In one embodiment, R^(4b) is HO₂CCH₂—.

In one embodiment, R^(4b) is (1-6C alkyl)NH(CO)—. Examples includeCH₃NHC(O)—, CH₃CH₂NHC(O)—, CH₃CH₂CH₂NHC(O)—, and (CH₃)₂CHNHC(O)—. In oneembodiment, R⁴ is CH₃NHC(O)—.

In one embodiment, R^(4b) is selected from H, methyl, —C(O)CH₃,methylsulfonyl, —C(O)CH₂OH, —CH₂COOH and —C(O)NHCH₂CH₃.

In one embodiment, Z is *—OC(═O)—.

In one embodiment of Formula I, ring B is ring B-1:

where R⁵ and R⁶ are independently H, halogen, OH, (1-6C)alkyl orhydroxy(1-6C)alkyl.

In one embodiment, R⁵ and R⁶ are independently H, F, OH, (1-6C)alkyl orhydroxy(1-6C)alkyl. In one embodiment, R⁵ is H and R⁶ is H, F, OH,(1-6C)alkyl or hydroxy(1-6C)alkyl.

In one embodiment, R⁵ and R⁶ are independently H, F, OH, (1-3C)alkyl orhydroxy(1-3C)alkyl. In one embodiment, R⁵ is hydrogen and R⁶ is H, F,OH, (1-3C)alkyl or hydroxy(1-3C)alkyl.

In one embodiment, R⁵ and R⁶ are independently H, F, OH, methyl, ethyl,HOCH₂— or HOCH₂CH₇—. In one embodiment, R⁵ is hydrogen and R⁶ is H, F,OH, methyl, ethyl, HOCH₂— or HOCH₂CH₂—.

In one embodiment, R⁵ and R⁶ are independently H, F, or methyl. In oneembodiment, R⁵ is H and R⁶ is H, F, or methyl.

In one embodiment, R⁵ is H and R⁶ is F.

In one embodiment, R⁵ is H and R⁶ is methyl.

In one embodiment, R⁵ and R⁶ are both H.

In one embodiment, R⁵ and R⁶ are both F.

In one embodiment, R⁵ and R⁶ are both methyl.

In one embodiment, ring B is ring B-1 which is optionally substitutedwith one or two substituents independently selected from OH and F,provided that two OH substituents are not on the same ring carbon atom.

Particular examples of ring B when represented by ring B-1 include thestructures:

In one embodiment of Formula I, ring B is ring B-2 having the formula:

In one embodiment, m is 0.

In one embodiment, m is 1.

In one embodiment, m is 2.

One embodiment of this invention provides compounds of the generalFormula I or pharmaceutically acceptable salts or solvates thereof,wherein:

ring B is ring B-1:

ring A is selected from rings A-1, A-2 and A-3 having the structures:

wherein the wavy line labeled 1 indicates the point of attachment ofring A to the pyrrolidine ring of Formula I and the wavy line labeled 2indicates the point of attachment of ring A to W;

X is N or CH;

Y is H or F;

R¹ is H, (1-3C)alkoxy or halogen;

W is O, NH or CH₂, wherein when ring A is A-2, then W is CH₂;

m is 0, 1 or 2;

D is carbon;

R² and R^(2a) are independently H, F, (1-3C)alkyl or OH, provided thatR² and R^(2a) are not both OH;

R³ and R^(3a) are independently H, (1-3C)alkyl or hydroxy(1-3C)alkyl;

or R² and R³ are absent and R^(2a) and R^(3a) together with the atoms towhich they are attached form a bivalent 5-6 membered heteroaryl ringhaving 1-2 ring nitrogen atoms;

Z is *—NR^(4a)C(═O)—, *—ONHC(═O)—, *—NR^(4b)CH₂— or *—OC(═O)—, whereinthe asterisk indicates the point of attachment of Z to the carbonbearing R³;

R^(4a) is H, (1-6C)alkyl, fluoro(1-6C)alkyl, difluoro(1-6C)alkyl,trifluoro(1-6C)alkyl, hydroxy(1-6C alkyl) or dihydroxy(2-6C alkyl);

R^(4b) is H, (1-6C)alkyl, fluoro(1-6C)alkyl, difluoro(1-6C)alkyl,trifluoro(1-6C)alkyl, hydroxy(1-6C alkyl), dihydroxy(2-6C alkyl), (1-6Calkyl)C(O)—, (3-6C cycloalkyl)C(O)—, Ar¹C(O)—, HOCH₂C(O)—, (1-6Calkyl)sulfonyl, (3-6C cycloalkyl)sulfonyl, Ar²(SO₂)—, HO₂CCH₂— or (1-6Calkyl)NH(CO)—;

Ar¹ is phenyl optionally substituted with one or more substituentsindependently selected from halogen, (1-6C)alkyl, and (1-6C)alkoxy;

Ar² is phenyl optionally substituted with one or more substituentsindependently selected from halogen, (1-6C)alkyl, and (1-6C)alkoxy; and

R⁵ and R⁶ are independently H, halogen, OH, (1-6C)alkyl orhydroxy(1-6C)alkyl.

One embodiment of this invention provides compounds of the generalFormula IA

or pharmaceutically acceptable salts or solvates thereof, wherein:

ring A is selected from rings A-1, A-2 and A-3 having the structures:

wherein the wavy line labeled 1 indicates the point of attachment ofring A to the pyrrolidine ring of Formula I and the wavy line labeled 2indicates the point of attachment of ring A to W;

X is N or CH;

Y is H or F;

R¹ is H, (1-3C)alkoxy or halogen;

W is O, NH or CH₂, wherein when ring A is A-2, then W is CH₂;

m is 0, 1 or 2;

R² and R^(2a) are independently H, F, or OH, provided that R² and R^(2a)are not both OH;

R³ is H, (1-3C)alkyl or hydroxy(1-3C)alkyl;

Z is *—NR^(4a)C(═O)—, *—ONHC(═O)—, *—NR^(4b)CH₂— or *—OC(═O)—, whereinthe asterisk indicates the point of attachment of Z to the carbonbearing R³;

R^(4a) is H, (1-6C)alkyl, fluoro(1-6C)alkyl, difluoro(1-6C)alkyl,trifluoro(1-6C)alkyl, hydroxy(1-6C alkyl) or dihydroxy(2-6C alkyl);

R^(4b) is H, (1-6C)alkyl, fluoro(1-6C)alkyl, difluoro(1-6C)alkyl,trifluoro(1-6C)alkyl, hydroxy(1-6C alkyl), dihydroxy(2-6C alkyl), (1-6Calkyl)C(O)—, (3-6C cycloalkyl)C(O)—, Ar¹C(O)—, HOCH₂C(O)—, (1-6Calkyl)sulfonyl, (3-6C cycloalkyl)sulfonyl, Ar²(SO₂)—, HO₂CCH₂— or (1-6Calkyl)NH(CO)—;

Ar¹ is phenyl optionally substituted with one or more substituentsindependently selected from halogen, (1-6C)alkyl, and (1-6C)alkoxy;

Ar² is phenyl optionally substituted with one or more substituentsindependently selected from halogen, (1-6C)alkyl, and (1-6C)alkoxy; and

R⁵ and R⁶ are independently H, halogen, OH, (1-6C)alkyl orhydroxy(1-6C)alkyl.

In one embodiment, Formula IA includes compounds wherein:

ring A is ring A-1 represented by the structure

wherein the wavy line labeled 1 indicates the point of attachment ofring A to the pyrrolidine ring of Formula I and the wavy line labeled 2indicates the point of attachment of ring A to W;

ring B is ring B-1 represented by the structure:

wherein the wavy line labeled 3 indicates the point of attachment toring A and the wavy line labeled 4 indicates the point of attachment tothe pyrazolo[1,5-a]pyrimidine ring of Formula I;

X is N or CH;

Y is H or F;

R¹ is H, (1-3C)alkyl, (1-3C)alkoxy or halogen;

W is O or NH;

m is 0, 1 or 2;

R² and R^(2a) are independently H, F, or OH, provided that R² and R^(2a)are not both OH;

R³ is H, (1-3C)alkyl or hydroxy(1-3C)alkyl;

Z is *—NR^(4a)C(═O)—, *—ONHC(═O)—, or *—OC(═O)—, wherein the asteriskindicates the point of attachment to the carbon bearing R¹;

R^(4a) is H, (1-6C)alkyl, fluoro(1-6C)alkyl, difluoro(1-6C)alkyl,trifluoro (1-6C)alkyl, hydroxy(1-6C alkyl) or dihydroxy(1-6C alkyl); and

R⁵ and R⁶ are independently H, halogen, OH, (1-6C)alkyl orhydroxy(1-6C)alkyl.

In one embodiment, X is N. In one embodiment, X is CH.

In one embodiment, Formula IA includes compounds wherein:

ring A is ring A-2 represented by the structure

wherein the wavy line labeled 1 indicates the point of attachment ofring A to the pyrrolidine ring of Formula I and the wavy line labeled 2indicates the point of attachment of ring A to W;

ring B is ring B-1 represented by the structure:

wherein the wavy line labeled 3 indicates the point of attachment toring A and the wavy line labeled 4 indicates the point of attachment tothe pyrazolo[1,5-a]pyrimidine ring of Formula I;

Y is H or F;

R¹ is H, (1-3C)alkyl, (1-3C)alkoxy or halogen;

m is 0, 1 or 2;

W is CH₂;

m is 0, 1 or 2;

R² and R^(2a) are independently H, F, or OH, provided that R² and R^(2a)are not both OH;

R³ is H, (1-3C)alkyl or hydroxy(1-3C)alkyl;

Z is *—NR^(4a)C(═O)—, wherein the asterisk indicates the point ofattachment to the carbon bearing R³;

R^(4a) is H, (1-6C)alkyl, fluoro(1-6C)alkyl, difluoro(1-6C)alkyl,trifluoro (1-6C)alkyl, hydroxy(1-6C alkyl) or dihydroxy(1-6C alkyl); and

R⁵ and R⁶ are independently H, halogen, OH, (1-6C)alkyl orhydroxy(1-6C)alkyl.

In one embodiment, Formula IA includes compounds wherein:

ring A is ring A-3 represented by the structure

wherein the wavy line labeled 1 indicates the point of attachment ofring A to the pyrrolidine ring of Formula I and the wavy line labeled 2indicates the point of attachment of ring A to W;

ring B is ring B-1 represented by the structure:

wherein the wavy line labeled 3 indicates the point of attachment toring A and the wavy line labeled 4 indicates the point of attachment tothe pyrazolo[1,5-a]pyrimidine ring of Formula I;

Y is H or F;

R¹ is H, (1-3C)alkyl, (1-3C)alkoxy or halogen;

W is O;

m is 0, 1 or 2;

R² and R^(2a) are independently H, F, or OH, provided that R² and R^(2a)are not both OH;

R³ is H, (1-3C)alkyl or hydroxy(1-3C)alkyl;

Z is *—OC(═O)— or *—NR^(4a)C(═O)—, wherein the asterisk indicates thepoint of attachment to the carbon bearing R³;

R^(4a) is H, (1-6C)alkyl, fluoro(1-6C)alkyl, difluoro(1-6C)alkyl,trifluoro (1-6C)alkyl, hydroxy(1-6C alkyl) or dihydroxy(1-6C alkyl); and

R⁵ and R⁶ are independently H, halogen, OH, (1-6C)alkyl orhydroxy(1-6C)alkyl.

In one embodiment, Formula IA includes compounds wherein:

ring A is ring A-1 represented by the structure

wherein the wavy line labeled 1 indicates the point of attachment ofring A to the pyrrolidine ring of Formula I and the wavy line labeled 2indicates the point of attachment of ring A to W;

ring B is ring B-1 represented by the structure:

wherein the wavy line labeled 3 indicates the point of attachment toring A and the wavy line labeled 4 indicates the point of attachment tothe pyrazolo[1,5-a]pyrimidine ring of Formula I;

X is N or CH;

Y is H or F;

R¹ is H, (1-3C)alkyl, (1-3C)alkoxy or halogen;

W is O;

m is 0, 1 or 2;

R² and R^(2a) are independently H, F, or OH, provided that R² and R^(2a)are not both OH;

R³ is H, (1-3C)alkyl or hydroxy(1-3C)alkyl;

Z is *—NR^(4b)CH₂—, wherein the asterisk indicates the point ofattachment to the carbon bearing R³;

R^(4b) is H, (1-6C)alkyl, fluoro(1-6C)alkyl, difluoro(1-6C)alkyl,trifluoro(1-6C)alkyl (1-6C alkyl)C(O)—, (3-6C cycloalkyl)C(O)—,Ar^(i)C(O)—, HOCH₂C(O)—, (1-6C alkyl)sulfonyl, (3-6Ccycloalkyl)sulfonyl, Ar²(SO₂)—, HO₂CCH₂— or (1-6C alkyl)NH(CO)—;

Ar¹ is phenyl optionally substituted with one or more substituentsindependently selected from halogen, (1-6C)alkyl, and (1-6C)alkoxy;

Ar² is phenyl optionally substituted with one or more substituentsindependently selected from halogen, (1-6C)alkyl, and (1-6C)alkoxy; and

R⁵ and R⁶ are independently H, halogen, OH, (1-6C)alkyl orhydroxy(1-6C)alkyl.

It will be appreciated that certain compounds according to the inventionmay contain one or more centers of asymmetry and may therefore beprepared and isolated as a mixture of isomers such as a racemic ordiastereomeric mixture, or in an enantiomerically or diastereomericallypure form. It is intended that all stereoisomeric forms of the compoundsof the invention, including but not limited to, diastereomers,enantiomers and atropisomers, as well as mixtures thereof such asracemic mixtures, form part of the present invention.

In one embodiment, compounds of the general Formula I wherein Ring B isring B-1 have the absolute configuration of FIG. 1-a:

In one embodiment, compounds of the general Formula I wherein Ring B isring B-1 have the absolute configuration of FIG. 1-b:

In the structures shown herein, where the stereochemistry of anyparticular chiral atom is not specified, then all stereoisomers arecontemplated and included as the compounds of the invention. Wherestereochemistry is specified by a solid wedge or dashed linerepresenting a particular configuration, then that stereoisomer is sospecified and defined.

The terms “(1-3C)alkyl” and “(1-6C)alkyl” as used herein refer tosaturated linear or branched-chain monovalent hydrocarbon radicals ofone to three carbon atoms and one to six carbon atoms, respectively.Examples include, but are not limited to, methyl, ethyl, 1-propyl,isopropyl, 1-butyl, isobutyl, sec-butyl, tert-butyl, 2-methyl-2-propyl,pentyl, and hexyl.

The term “fluoro(1-6C)alkyl” as used herein refers to saturated linearor branched-chain monovalent hydrocarbon radicals of one to six carbonatoms as defined herein, wherein one of the hydrogens is replaced by afluorine atom.

The term “difluoro(1-6C)alkyl” as used herein refers to saturated linearor branched-chain monovalent hydrocarbon radicals of one to six carbonatoms as defined herein, wherein two of the hydrogens are replaced byfluorine atoms.

The term “trifluoro(1-6C)alkyl” as used herein refers to saturatedlinear or branched-chain monovalent hydrocarbon radicals of one to sixcarbon atoms as defined herein, wherein three of the hydrogens arereplaced by fluorine atoms.

The term “hydroxy(1-6Calkyl) as used herein refers to saturated linearor branched-chain monovalent hydrocarbon radicals of one to six carbonatoms, wherein one of the hydrogens is replaced by a hydroxy (OH) group.

The term “dihydroxy(1-6Calkyl) as used herein refers to saturated linearor branched-chain monovalent hydrocarbon radicals of one to six carbonatoms as defined herein, wherein two of the hydrogens are replaced byhydroxy (OH) groups, provided the hydroxy groups are not on the samecarbon atom.

The term “(1-6C alkyl)sulfonyl” as used herein refers to a (1-6Calkyl)SO₂— group, wherein the radical is on the sulfur atom and the(1-6C alkyl) portion is as defined above. Examples includemethylsulfonyl (CH₃SO₂—) and ethylsulfonyl (CH₃CH₂SO₂—).

The term “(3-6C cycloalkyl)sulfonyl” as used herein refers to a (3-6Ccycloalkyl)SO₂— group, wherein the radical is on the sulfur atom. Anexample is cyclopropylsulfonyl.

The terms “(1-4C)alkoxy” and “(1-6C)alkoxy”, as used herein refer tosaturated linear or branched-chain monovalent alkoxy radicals of one tofour carbon atoms or one to six carbon atoms, respectively, wherein theradical is on the oxygen atom. Examples include methoxy, ethoxy,propoxy, isopropoxy, and butoxy.

The term “halogen” includes fluoro, chloro, bromo and iodo.

It will also be appreciated that certain compounds of Formula I may beused as intermediates for the preparation of further compounds ofFormula I.

The compounds of Formula I include salts thereof. In certainembodiments, the salts are pharmaceutically acceptable salts. Inaddition, the compounds of Formula I include other salts of suchcompounds which are not necessarily pharmaceutically acceptable salts,and which may be useful as intermediates for preparing and/or purifyingcompounds of Formula I and/or for separating enantiomers of compounds ofFormula I.

The term “pharmaceutically acceptable” indicates that the substance orcomposition is compatible chemically and/or toxicologically, with theother ingredients comprising a formulation, and/or the mammal beingtreated therewith.

It will further be appreciated that the compounds of Formula I and theirsalts may be isolated in the form of solvates, and accordingly that anysuch solvate is included within the scope of the present invention.

Compounds of the invention may also contain unnatural proportions ofatomic isotopes at one or more of the atoms that constitute suchcompounds. That is, an atom, in particular when mentioned in relation toa compound according to Formula I, comprises all isotopes and isotopicmixtures of that atom, either naturally occurring or syntheticallyproduced, either with natural abundance or in an isotopically enrichedform. For example, when hydrogen is mentioned, it is understood to referto ¹H, ²H, ³H or mixtures thereof; when carbon is mentioned, it isunderstood to refer to ¹¹C, ¹²C, ¹³C, ¹⁴C or mixtures thereof; whennitrogen is mentioned, it is understood to refer to ¹³N, ¹⁴N, ¹⁵N ormixtures thereof; when oxygen is mentioned, it is understood to refer to¹⁴O, ¹⁵O, ¹⁶O, ¹⁷O, ¹⁸O, or mixtures thereof; and when fluoro ismentioned, it is understood to refer to ¹⁸F, ¹⁹F or mixtures thereof.The compounds according to the invention therefore also comprisecompounds with one or more isotopes of one or more atom, and mixturesthereof, including radioactive compounds, wherein one or morenon-radioactive atoms has been replaced by one of its radioactiveenriched isotopes. Radiolabeled compounds are useful as therapeuticagents, e.g., cancer therapeutic agents, research reagents, e.g., assayreagents, and diagnostic agents, e.g., in vivo imaging agents. Allisotopic variations of the compounds of the present invention, whetherradioactive or not, are intended to be encompassed within the scope ofthe present invention.

The present invention further provides a process for the preparation ofa compound of Formula I or a salt thereof as defined herein whichcomprises:

(a) for a compound of Formula I wherein Z is *—NHC(═O)—, and ring A,ring B, W, D, R², R^(2a), R³, R^(3a) and m are as defined for Formula I,cyclizing a corresponding compound having the formula II

where P¹ is H or a carboxyl protecting group, in the presence of acoupling reagent and a base; or

(b) for a compound of Formula I wherein W is O, ring A is formula A-1:

X is N, and ring B, D, Z, Y, R¹, R², R^(2a), R³, R^(3a) and m are asdefined for Formula I, cyclizing a corresponding compound having theformula III

where n is 1, 2, 3 or 4 and L′ is a leaving group or atom, in thepresence of a base; or

(c) for a compound of Formula I wherein W is CH₂, ring A is formula A-2:

and ring B, Z, D, Y, R¹, R², R^(2a), R³, R^(3a) and m are as defined forFormula I, cyclizing a corresponding compound having the formula IV

where L² is a leaving group or atom, in the presence of a base; or

(d) for a compound of Formula I wherein Z is *—NHC(═O)—, and ring A,ring B, W, D, R², R^(2a), R³, R^(3a) and m are as defined for Formula I,cyclizing a corresponding compound having the formula V

in the presence of a base and a coupling reagent; or

(e) for a compound of Formula I wherein Z is *—NHCH₂—, and ring A, ringB, W, D, R², R^(2a), R³, R^(3a) and m are as defined for Formula I,cyclizing a corresponding compound having the formula VI

in the presence of a reducing agent; or

(f) for a compound of Formula I wherein Z is *—NHCH₂—, and ring A, ringB, W, D, R², R^(2a), R³, R^(3a) and m are as defined for Formula I,cyclizing a corresponding compound having the formula VII

in the presence of triphenylphosphine; or

(g) for a compound of Formula I wherein ring A, ring B, W, D, m, R²,R^(2a), R³, and R^(3a) are as defined for Formula I, Z is *—NR^(4b)CH₂—,and R^(4b) is (1-6C alkyl)C(O)—, (3-6C cycloalkyl)C(O)—, Ar¹C(O)—,HOCH₂C(O)—, (1-6C alkyl)sulfonyl, (3-6C cycloalkyl)sulfonyl, (1-6Calkyl)sulfonyl, (3-6C cycloalkyl)sulfonyl, or Ar²(SO₂)—, coupling acorresponding compound having the formula VIII

with a reagent having the formula (1-6C alkyl)C(O)-L³, (3-6Ccycloalkyl)C(O)-L³, Ar′C(O)-L³, HOCH₂C(O)-L³, (1-6C alkyl)(SO₂)-L³,(3-6C cycloalkyl)(SO₂)-L³, or Ar²(SO₂)-L³, respectively, where L³ is aleaving atom, in the presence of a base; or

(h) for a compound of Formula I wherein ring A, ring B, W, D, R²,R^(2a), R³, R^(3a) and m are as defined for Formula I, Z is*—NR^(4b)CH₂—, and R^(4b) is (1-6C alkyl)NH(CO)—, reacting a compoundhaving the formula VIII

with a reagent having the formula (1-6C alkyl)N═C═O in the presence of abase; or

(i) for a compound of Formula I wherein R² is F, R^(2a) is H, and ringA, ring B, Z, W, D, R³, R^(3a), and m are as defined for Formula I,reacting a corresponding compound having the formula IX

with a fluorination reagent;

(j) for a compound of Formula I wherein W is O, ring A is formula A-1,

X is CH, and Y, R¹, D, ring B, Z, R², R^(2a), R³ and m are as definedfor Formula I, cyclizing a corresponding compound having the formula X

where n is 1, 2, 3 or 4 and L¹ is a leaving group or atom, in thepresence of a base; and

optionally removing any protecting groups and optionally preparing asalt thereof.

In one embodiment of the above-described methods (a)-(j), ring B is ringB-1 having the structure:

D is carbon, R² and R^(2a) are independently H, F, (1-3C)alkyl or OH(provided that R² and R^(2a) are not both OH), R³ is H, (1-3C)alkyl orhydroxy(1-3C)alkyl., and ring A, W, m, Z, R⁵ and R⁶ are as defined forFormula I.

Referring to method (a), the cyclization may be performed usingconventional amide bond formation conditions, for example by treatingthe carboxylic acid with an activating agent, followed by addition ofthe amine in the presence of a base. Suitable activating agents includeEDCI, oxalyl chloride, thionyl chloride, HATU, and HOBt. Suitable basesinclude amine bases, for example triethylamine, diisopropylethylamine,pyridine, or excess ammonia. Suitable solvents include DCM, DCE, THF andDMF.

Referring to methods (b) and (c), the leaving atoms L¹ and L² may be,for example a halogen atom such as Br, Cl or I. Alternatively, L¹ and L²can be a leaving group, for example an arylsulfonyloxy group or analkylsulfonyloxy group, such as a mesylate or a tosylate group. Suitablebases include alkali metal carbonates, such as sodium carbonate,potassium carbonate or cesium carbonate. Convenient solvents includeaprotic solvents such as ethers (for example tetrahydrofuran orp-dioxane), DMF, or acetone. The reaction can be conveniently performedat elevated temperatures, for example 50-150° C., for example at 85° C.

Referring to method (d), suitable coupling reagents include HATU, HBTU,TBTU, DCC, DIEC, and any other amide coupling reagents well known topersons skilled in the art. Suitable bases include tertiary amine basessuch as DIEA and triethylamine. Convenient solvents include DMF, THF,DCM and DCE.

Referring to method (e), suitable reducing agents include Me₄N(OAc)₃BH,Na(OAc)₃BH and NaCNBH₃. Suitable solvents include neutral solvents suchas acetonitrile, THF and DCE. The reaction can be conveniently performedat ambient temperature.

Referring to method (f), in certain embodiments the triphenylphosphinereagent is used in the form of a polystyrene-bound PPh₃ resin (sold asPS—PPh₃ by Biotage Systems). The reaction is conveniently performed atambient temperature. Suitable solvents include neutral solvents, forexample DCM.

Referring to method (g), the leaving atom L³ may be a halogen, forexample Cl or Br. Suitable bases include tertiary amine bases such asdiisopropylethylamine and triethylamine. The reaction is convenientlyperformed at ambient temperature.

Referring to method (h), suitable bases include tertiary amine basessuch as DIEA and triethylamine. The reaction is conveniently performedat ambient temperature.

Referring to method (i), the fluorination reagent may be, for example,bis(2-methoxyethyl)amino-sulfur trifluoride (Deoxo-Fluor™) ordiethylaminosulfur trifluoride (DAST). Suitable solvents includedichloromethane, chloroform, dichloroethane, and toluene. The reactionis conveniently performed at ambient temperature.

Referring to method (j), base may be, for example, an alkali metalcarbonate, such as for example sodium carbonate, potassium carbonate orcesium carbonate. Convenient solvents include aprotic solvents such asethers (for example tetrahydrofuran or p-dioxane) or toluene. Thereaction can be conveniently performed at a temperature between ambienttemperature and reflux, for example at 85° C.

Amine groups in compounds described in any of the above methods may beprotected with any convenient amine protecting group, for example asdescribed in Greene & Wuts, eds., “Protecting Groups in OrganicSynthesis”, 2^(nd) ed. New York; John Wiley & Sons, Inc., 1991. Examplesof amine protecting groups include acyl and alkoxycarbonyl groups, suchas t-butoxycarbonyl (BOC), and [2-(trimethylsilyl)ethoxy]methyl (SEM).Likewise, carboxyl groups may be protected with any convenient carboxylprotecting group, for example as described in Greene & Wuts, eds.,“Protecting Groups in Organic Synthesis”, 2^(nd) ed. New York; JohnWiley & Sons, Inc., 1991. Examples of carboxyl protecting groups include(1-6C)alkyl groups, such as methyl, ethyl and t-butyl. Alcohol groupsmay be protected with any convenient alcohol protecting group, forexample as described in Greene & Wuts, eds., “Protecting Groups inOrganic Synthesis”, 2^(nd) ed. New York; John Wiley & Sons, Inc., 1991.Examples of alcohol protecting groups include benzyl, trityl, silylethers, and the like.

The compounds of the formulas II, III, IV, V, VI, VII, VIII, IX and Xare also believed to be novel and are provided as further aspects of theinvention.

The ability of compounds of the invention to act as TrkA inhibitors maybe demonstrated by the assays described in Examples A and B.

Certain compounds which are inhibitors of TrkA and/or TrkB may be usefulin the treatment of multiple types of pain including inflammatory pain,neuropathic pain, and pain associated with cancer, surgery, and bonefracture.

In one embodiment, compounds of Formula I are useful for treating pain,including chronic and acute pain, in a mammal.

Acute pain, as defined by the International Association for the Study ofPain, results from disease, inflammation, or injury to tissues. Thistype of pain generally comes on suddenly, for example, after trauma orsurgery, and may be accompanied by anxiety or stress. The cause canusually be diagnosed and treated, and the pain is confined to a givenperiod of time and severity. In some rare instances, it can becomechronic.

Chronic pain, as defined by the International Association for the Studyof Pain, is widely believed to represent disease itself. It can be mademuch worse by environmental and psychological factors. Chronic painpersists over a longer period than acute pain and is resistant to mostmedical treatments, generally over 3 months or more. It can and oftendoes cause severe problems for patients.

Compounds of Formula I are also useful for treating cancer in a mammal.Particular examples include neuroblastoma, ovarian, pancreatic,colorectal and prostate cancer.

Compounds of Formula I are also useful for treating inflammation in amammal.

Compounds of Formula I are also useful for treating certain infectiousdiseases in a mammal, such as Trypanosoma cruzi infection.

Compounds of Formula I may also be used to treat neurodegenerativediseases in a mammal. Examples of neurodegenerative disease includedemyelination and dysmyelination. Additional examples ofneurodegenerative diseases include multiple sclerosis, Parkinson'sdisease and Alzheimer's disease.

In addition, compounds of Formula I may also be used to treatinterstitial cystitis (IC), painful bladder syndrome (PBS), urinaryincontinence, asthma, anorexia, atopic dermatitis, and psoriasis in amammal.

Accordingly, another embodiment of this invention provides a method oftreating or preventing pain in a mammal, comprising administering tosaid mammal one or more compounds of Formula I or a pharmaceuticallyacceptable salt thereof in an amount effective to treat or prevent saidpain. In one embodiment, the pain is chronic pain. In one embodiment,the pain is acute pain. In one embodiment, the pain is inflammatorypain. In one embodiment, the pain is neuropathic pain. In oneembodiment, the pain is pain associated with cancer. In one embodiment,the pain is pain associated with surgery. In one embodiment, the pain ispain associated with bone fracture. In one embodiment, the methodcomprises a method of treating said pain in a mammal. In one embodiment,the method comprises a method of preventing said pain in a mammal.

Another embodiment of this invention provides a method of treating orpreventing inflammation in a mammal, comprising administering to saidmammal one or more compounds of Formula I or a pharmaceuticallyacceptable salt thereof in an amount effective to treat or prevent saidinflammation. In one embodiment, the method comprises a method oftreating said inflammation in a mammal. In one embodiment, the methodcomprises a method of preventing said inflammation in a mammal.

Another embodiment of this invention provides a method of treating orpreventing a neurodegenerative disease in a mammal, comprisingadministering to said mammal one or more compounds of Formula I or apharmaceutically acceptable salt thereof in an amount effective to treator prevent said neurodegenerative disease. In one embodiment, theneurodegenerative disease is demyelination. In one embodiment, theneurodegenerative disease is dysmyelination. In one embodiment, theneurodegenerative disease is multiple sclerosis. In one embodiment, theneurodegenerative disease is Parkinson's disease. In one embodiment, theneurodegenerative disease is Alzheimer's disease. Another embodiment ofthis invention provides a method of treating or preventing infectiousdiseases in a mammal, comprising administering to said mammal one ormore compounds of Formula I or a pharmaceutically acceptable saltthereof in an amount effective to treat or prevent said infectiousdisease. In one embodiment, the infectious disease is Trypanosoma cruziinfection. In one embodiment, the method comprises a method of treatingsaid neurodegenerative disease in a mammal. In one embodiment, themethod comprises a method of preventing said neurodegenerative diseasein a mammal.

Another embodiment of this invention provides a method of treating orpreventing cancer in a mammal, comprising administering to said mammalone or more compounds of Formula I or a pharmaceutically acceptable saltthereof in an amount effective to treat or prevent said cancer. In oneembodiment, the cancer is neuroblastoma. In one embodiment, the canceris ovarian cancer. In one embodiment, the cancer is pancreatic cancer.In one embodiment, the cancer is colorectal cancer. In one embodiment,the cancer is prostate cancer. In one embodiment, the method comprises amethod of treating said cancer in a mammal. In one embodiment, themethod comprises a method of preventing said cancer in a mammal.

Compounds of Formula I may be administered alone as a sole therapy orcan be administered in addition with one or more other substances and/ortreatments that work by the same or a different mechanism of action.Examples include anti-inflammatory compounds, steroids (e.g.,dexamethasone, cortisone and fluticasone), analgesics such as NSAIDs(e.g., aspirin, ibuprofen, indomethacin, and ketoprofen), and opioids(such as morphine), and chemotherapeutic agents. These agents may beadministered with one or more compounds of Formula I as part of the sameor separate dosage forms, via the same or different routes ofadministration, and on the same or different administration schedulesaccording to standard pharmaceutical practice known to one skilled inthe art.

In the field of medical oncology it is normal practice to use acombination of different forms of treatment to treat each patient withcancer. In medical oncology the other component(s) of such conjointtreatment in addition to compositions of the present invention may be,for example, surgery, radiotherapy, chemotherapy, signal transductioninhibitors and/or monoclonal antibodies.

Accordingly, the compounds of Formula I may be administered incombination with one or more agents selected from mitotic inhibitors,alkylating agents, anti-metabolites, antisense DNA or RNA, intercalatingantibiotics, growth factor inhibitors, signal transduction inhibitors,cell cycle inhibitors, enzyme inhibitors, retinoid receptor modulators,proteasome inhibitors, topoisomerase inhibitors, biological responsemodifiers, anti-hormones, angiogenesis inhibitors, cytostatic agentsanti-androgens, targeted antibodies, HMG-CoA reductase inhibitors, andprenyl-protein transferase inhibitors. These agents may be administeredwith one or more compounds of Formula I as part of the same or separatedosage forms, via the same or different routes of administration, and onthe same or different administration schedules according to standardpharmaceutical practice known to one skilled in the art.

As used herein, terms “treat” or “treatment” refer to therapeutic,prophylactic, palliative or preventative measures. Beneficial or desiredclinical results include, but are not limited to, alleviation ofsymptoms, diminishment of extent of disease, stabilized (i.e., notworsening) state of disease, delay or slowing of disease progression,amelioration or palliation of the disease state, and remission (whetherpartial or total), whether detectable or undetectable. “Treatment” canalso mean prolonging survival as compared to expected survival if notreceiving treatment. Those in need of treatment include those alreadywith the condition or disorder, as well as those prone to have thecondition or disorder or those in which the condition or disorder is tobe prevented.

In one embodiment, the terms “treatment” or “treating” as used herein,mean an alleviation, in whole or in part, of symptoms associated with adisorder or condition as described herein (e.g., multiple types of painincluding inflammatory pain, neuropathic pain, and pain associated withcancer, surgery, and bone fracture), or slowing, or halting of furtherprogression or worsening of those symptoms.

In one embodiment, the term “preventing” as used herein means theprevention of the onset, recurrence or spread, in whole or in part, ofthe disease or condition as described herein (e.g., multiple types ofpain including inflammatory pain, neuropathic pain, and pain associatedwith cancer, surgery, and bone fracture), or a symptom thereof.

The terms “effective amount” and “therapeutically effective amount”refer to an amount of compound that, when administered to a mammal inneed of such treatment, is sufficient to (i) treat or prevent aparticular disease, condition, or disorder, (ii) attenuate, ameliorate,or eliminate one or more symptoms of the particular disease, condition,or disorder, or (iii) prevent or delay the onset of one or more symptomsof the particular disease, condition, or disorder described herein. Theamount of a compound of Formula I that will correspond to such an amountwill vary depending upon factors such as the particular compound,disease condition and its severity, the identity (e.g., weight) of themammal in need of treatment, but can nevertheless be routinelydetermined by one skilled in the art.

As used herein, the term “mammal” refers to a warm-blooded animal thathas or is at risk of developing a disease described herein and includes,but is not limited to, guinea pigs, dogs, cats, rats, mice, hamsters,and primates, including humans.

Compounds of the invention may be administered by any convenient route,e.g. into the gastrointestinal tract (e.g. rectally or orally), thenose, lungs, musculature or vasculature, or transdermally or dermally.Compounds may be administered in any convenient administrative form,e.g. tablets, powders, capsules, solutions, dispersions, suspensions,syrups, sprays, suppositories, gels, emulsions, patches etc. Suchcompositions may contain components conventional in pharmaceuticalpreparations, e.g. diluents, carriers, pH modifiers, sweeteners, bulkingagents, and further active agents. If parenteral administration isdesired, the compositions will be sterile and in a solution orsuspension form suitable for injection or infusion. Such compositionsform a further aspect of the invention.

The present invention further provides a pharmaceutical composition,which comprises a compound of Formula I or a pharmaceutically acceptablesalt thereof, as defined hereinabove. In one embodiment, thepharmaceutical composition includes the compound of Formula I togetherwith a pharmaceutically acceptable diluent or carrier.

The present invention further provides a compound of Formula I or apharmaceutically acceptable salt thereof, for use in therapy.

In one embodiment, the invention provides a compound of Formula I or apharmaceutically acceptable salt thereof, for use in the treatment ofpain in a mammal. In one embodiment, the pain is chronic pain. In oneembodiment, the pain is acute pain. In one embodiment, the pain isinflammatory pain. In one embodiment, the pain is neuropathic pain. Inone embodiment, the pain is pain associated with cancer. In oneembodiment, the pain is pain associated with surgery. In one embodiment,the pain is pain associated with bone fracture.

In one embodiment, the invention provides a compound of Formula I or apharmaceutically acceptable salt thereof, for use in the treatment ofinflammation in a mammal.

According to a further aspect, the present invention provides a compoundof Formula I or a pharmaceutically acceptable salt thereof, for use inthe treatment of a neurodegenerative disease in a mammal. In oneembodiment, the neurodegenerative disease is demyelination. In oneembodiment, the neurodegenerative disease is dysmyelination. In oneembodiment, the neurodegenerative disease is multiple sclerosis. In oneembodiment, the neurodegenerative disease is Parkinson's disease. In oneembodiment, the neurodegenerative disease is Alzheimer's disease.

In one embodiment, the invention provides a compound of Formula I or apharmaceutically acceptable salt thereof, for use in the treatment ofinfectious diseases in a mammal. In one embodiment, the infectiousdisease is Trypanosoma cruzi infection.

In one embodiment, the invention provides a compound of Formula I or apharmaceutically acceptable salt thereof, for use in the treatment ofcancer in a mammal. In one embodiment, the cancer is neuroblastoma. Inone embodiment, the cancer is ovarian cancer. In one embodiment, thecancer is pancreatic cancer. In one embodiment, the cancer is colorectalcancer. In one embodiment, the cancer is prostate cancer.

Another embodiment of the present invention provides the use of acompound of Formula I, or a pharmaceutically acceptable salt thereof, inthe manufacture of a medicament for the treatment of pain in a mammal.

Another embodiment of the present invention provides the use of acompound of Formula I, or a pharmaccutically acceptable salt thereof, inthe manufacture of a medicament for the treatment of inflammation in amammal.

Another embodiment of the present invention provides the use of acompound of Formula I, or a pharmaceutically acceptable salt thereof, inthe manufacture of a medicament for the treatment of a neurodegenerativedisease in a mammal.

Another embodiment of the present invention provides the use of acompound of Formula I, or a pharmaceutically acceptable salt thereof, inthe manufacture of a medicament for the treatment of infectious diseasesin a mammal.

Another embodiment of the present invention provides the use of acompound of Formula I, or a pharmaceutically acceptable salt thereof, inthe manufacture of a medicament for the treatment of cancer in a mammal.

EXAMPLES

The following examples illustrate the invention. In the examplesdescribed below, unless otherwise indicated all temperatures are setforth in degrees Celsius. Reagents were purchased from commercialsuppliers such as Aldrich Chemical Company, Lancaster, Alfa, Aesar, TCI,Maybridge, Asta Tech, or other suitable suppliers, and were used withoutfurther purification unless otherwise indicated. THF, DCM, toluene, DMFand dioxane were purchased from Aldrich in Sure/Sealim bottles and usedas received.

The reactions set forth below were done generally under a positivepressure of nitrogen or argon or with a drying tube (unless otherwisestated) in anhydrous solvents, and the reaction flasks were typicallyfitted with rubber septa for the introduction of substrates and reagentsvia syringe. Glassware was oven dried and/or heat dried or dried under astream of dry nitrogen.

Column chromatography was done on a Biotage system (Manufacturer: DyaxCorporation) having a silica gel or C-18 reverse phase column, or on asilica SepPak cartridge (Waters), or using conventional flash columnchromatography on silica gel, unless otherwise specified.

Abbreviations used herein have the following meanings:

CAN acetonitrile APCI Atmospheric Pressure Chemical Ionization BINAP2,2′-bis(diphenylphosphino)-1,1′-binaphthyl Boc tert-butoxycarbonylBoc₂O Di-tert-butyl dicarbonate BOP(Benzotriazol-1-yloxy)tris(dimethylamino)phosphonium hexafluorophosphateCDI carbonyl diimidazole DCC N,N′-dicyclohexylcarbodiimide DCEDichloroethane DCM Dichloromethane DIEA Diisopropylethylamine DIEC1-(3-dimethylaminopropyl)-3-ethylcarboiimide DIPHOS1,2-Bis(Diphenylphosphino)ethane DMF N,N-Dimethylformamide DMSOdimethylsulfoxide EDCI 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide)HATU (2-(7-Aza-1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluroniumhexafluorophosphate) HOBt Hydroxybenzotriazole IPA Isopropyl alcoholMTBE tert-butyl-methylether Pd(PPh₃)₄Tetrakis(triphenylphosphine)palladium (0) Pd₂dba₃Tris(dibenzylideneacetone)dipalladium (0) PS-PPh₃ polystyrene-bound PPh₃resin TEA triethylamine TFA Trifluoroacetic acid THF Tetrahydrofuran TLCThin layer chromatography

Biological Assays Example A TrkA ELISA Assay

An enzyme-linked immunosorbant assay (ELISA) was used to assess TrkAkinase activity in the presence of inhibitors Immulon 4HBX 384-wellmicrotiter plates (Thermo part #8755) were coated with a 0.025 mg/mLsolution of poly (Glu, Ala, Tyr; 6:3:1; Sigma P3899). Variousconcentrations of test compound, 2.5 nM TrkA (Invitrogen Corp.,histidine-tagged recombinant human TrkA, cytoplasmic domain), and 500 μMATP were incubated for 25 minutes at ambient temperature in the coatedplates while shaking. The assay buffer consisted of 25 mM MOPS pH 7.5,0.005% (v/v) Triton X-100 and 5 mM MgCl₂. The reaction mixture wasremoved from the plate by washing with PBS containing 0.1% (v/v) Tween20. The phosphorylated reaction product was detected using 0.2 μg/mL ofa phosphotyrosine specific monoclonal antibody (clone PY20) conjugatedto horseradish peroxidase in conjunction with the TMB PeroxidaseSubstrate System (KPL). After the addition of 1M phosphoric acid, thechromogenic substrate color intensity was quantitated via absorbance at450 nm. IC₅₀ values were calculated using either a 4 or 5-parameterlogistic curve fit.

Table 1 provides averaged IC₅₀ values for compounds of the inventionwhen tested in this assay. In Table 1, the letter “A” designates an IC₅₀value between about 1 and 100 nM, and the letter “B” designates an IC₅₀value >100 nM and <3000 nM.

Example B TrkA Binding Assay

The ability of a compound to bind to TrkA was measured by Invitrogen'sLanthaScreen™ Eu Kinase Binding Assay. In this assay, His-taggedrecombinant human TrkA (cytoplasmic domain) from Invitrogen is incubatedwith Invitrogen's Alexa-Fluor® Tracer 236, biotinylated anti-His, andeuropium-labeled Streptavidin, compound (2% DMSO final) in buffer (25 mMMOPS, pH 7.5, 5 mM MgCl2, 0.005% Triton X-100). After a 60-minuteincubation at 22° C., the reaction was measured using the EnVision viaTR-FRET dual wavelength detection, and the POC was calculated from theemission ratio. The compound dose response data was fit to a 4-parameterlogistic model and IC₅₀ was defined as the concentration of compound at50 POC.

Table 1 provides averaged IC₅₀ values for compounds of the inventionwhen tested in this assay. In Table 1, the letter “A” designates an IC₅₀value between about 1 and 100 nM, and the letter “B” designates an IC₅₀value >100 nM and <3000 nM.

TABLE 1 TrkA Elisa Enzyme Assay TrkA Binding Assay Example No. IC₅₀ IC₅₀ 1 A A  2 A A  3 A A  4 A A  5 A A  6 A A  7 A A  8 A A  9 A A 10 A A 11A A 12 A 13 A 14 A 15 B 16 A A 17 A 18 A A 19 A A 20 A A 21 B 22 A 23 AA 24 B 25 A 26 B B 27 A 28 A 29 A 30 A 31 B B 32 A 33 A 34 A 35 A 36 A37 A 38 A 39 A 40 A 41 A 41-B B¹ 42 A 42-B B¹ 43 A 43-B B¹ 44 A 44-B A¹45 A¹ Diastereomer 1 45 A¹ Diastereomer 2 ¹Compound may have beenIsolated along with the enantiomer and/or one or more diastereomers,which additional isomer(s) were believed to make up ≦1.5% of the totalamount isolated.

Preparation A

(R)-5-fluoro-2-methoxy-3-(pyrrolidin-2-yl)pyridine Step A: Preparationof (R)-tert-butyl2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidine-1-carboxylate

A solution of tert-butyl pyrrolidine-1-carboxylate (4.09 mL, 23.4 mmol)and (−)-sparteine (6.44 mL, 28.0 mmol) in MTBE (50 mL) was cooled to−78° C. and sec-BuLi (20 mL, 28.0 mmol, 1.4 M in cyclohexane) wasintroduced drop-wise by cannula, keeping the internal temperature under−78° C. The resulting solution was stirred for 3 hours at −78° C.,followed by addition of a solution of ZnCl₂ (21.0 mL, 21.0 mmol, 1M inEt₂O) drop-wise with rapid stirring, keeping the internal temperaturebelow −65° C. The resulting light suspension was stirred at −78° C. for10 minutes and then warmed to ambient temperature. The resulting mixturewas sequentially charged with 3-bromo-5-fluoro-2-methoxypyridine (5.05g, 24.5 mmol), Pd(OAc)₂ (0.262 g, 1.17 mmol) and t-Bu₃P—HBF₄ (0.407 g,1.40 mmol) in one portion. After stirring overnight at ambienttemperature, concentrated NH₄OH (1 mL) was added and the reaction wasstirred for 1 hour. The resulting slurry was filtered through Celite®and washed with Et₂O. The organic layer was filtered and concentrated,and the crude product was purified by silica column chromatography,eluting with 5% EtOAc/hexanes to give product (R)-tert-butyl2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidine-1-carboxylate as yellowoil (4.34 g, 63% yield).

Step B: Preparation of (R)-5-fluoro-2-methoxy-3-pyrrolidin-2-yl)pyridine

A DCM (12 mL) solution of TFA (11.3 mL, 146 mmol) was added to(R)-tert-butyl2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidine-1-carboxylate (4.33 g,14.6 mmol) and stirred at ambient temperature for 1 hour. The reactionwas then concentrated, taken up in EtOAc, then washed with NaHCO₃ andbrine. The organic phase was dried (MgSO₄), filtered, and concentrated,and the crude material was purified by silica column chromatographyeluting with a 1-2% 7 N NH₃-MeOH/DCM to afford(R)-5-fluoro-2-methoxy-3-(pyrrolidin-2-yl)pyridine as a liquid (1.40 g,49% yield).

The Enantiomeric Excess (ee %) of(R)-5-fluoro-2-methoxy-3-(pyrrolidin-2-yl)pyridine was Determined asFollows

To a propan-2-ol solution of small amount of((R)-5-fluoro-2-methoxy-3-(pyrrolidin-2-yl)pyridine was added excessN-(2,4-dinitro-5-fluorophenyl)-L-alanine amide (FDAA, Marfey's reagent).The mixture was heated to reflux for approximately two minutes. Aftercooling to ambient temperature, the reaction mixture was diluted withacetonitrile and analyzed by HPLC (YMC ODS-AQ 4.6×50 mm 3 μm 120 Åcolumn; mobile phase: 5-95% solvent B in A; solvent A: H₂O/1% IPA/10 mMammonium acetate, and solvent B: ACN/1% IPA/10 mM ammonium acetate; flowrate: 2 mL/min). The enantiomeric excess was determined from the peakareas of the two diastereomeric derivatives formed. The ee % of theproduct was determined to be >93%.

Preparation B

(R)-5-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylicacid Step A: Preparation of ethyl5-hydroxypyrazolo[1,5-a]pyrimidine-3-carboxylate

To a mixture of ethyl 3-amino-1H-pyrazole-4-carboxylate (25.0 g, 161mmol) and (E)-ethyl 3-ethoxyacrylate (35.8 ml, 242 mmol) in DMF (537 mL)was added cesium carbonate (78.7 g, 242 mmol), and the reaction washeated at 110° C. for 15 hours. After cooling to ambient temperature thereaction was acidified with acetic acid to pH 4. The resultingprecipitate was filtered, washed with water and EtOAc, to provide theproduct as a white solid. To recover additional product, the filtratewas concentrated, diluted with EtOAc (500 mL) and washed with H₂O (5×200mL). The resulting precipitate in the EtOAc layer was filtered andwashed with water and EtOAc to obtain a second batch product. The twobatches of product were combined and dried under reduced pressure toafford ethyl 5-hydroxypyrazolo[1,5-a]pyrimidine-3-carboxylate as a whitesolid (33.3 g, 100% yield). MS (apci) m/z=206.2 (M−H).

Step B: Preparation of ethyl5-chloropyrazolo[1,5-a]pyrimidine-3-carboxylate

Ethyl 5-hydroxypyrazolo[1,5-a]pyrimidine-3-carboxylate (22.7 g, 110mmol) was suspended in phosphoryl trichloride (100 mL) and heated toreflux. After heating for 2 hours, the reaction mixture was cooled andconcentrated to remove excess POCl₃. The residue was diluted in DCM (100mL) and slowly added to a flask containing ice water. The mixture wasseparated and the aqueous layer was extracted with DCM (2×200 mL). Thecombined organic layers were dried (MgSO₄), filtered and concentrated toafford ethyl 5-chloropyrazolo[1,5-a]pyrimidine-3-carboxylate as apale-yellow solid (24.2 g, 97.6% yield). MS (apci) m/z=225.9 (M+H).

Step C: Preparation of (R)-ethyl5-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate

A mixture of ethyl 5-chloropyrazolo[1,5-a]pyrimidine-3-carboxylate (0.75g, 3.32 mmol), (R)-5-fluoro-2-methoxy-3-(pyrrolidin-2-yl)pyridine(Preparation A, 0.984 g, 3.66 mmol), DIEA (2.32 mL, 13.3 mmol) andn-butanol (1.11 mL) was sealed in a pressure tube and heated at 90° C.for 48 hours. The reaction mixture was diluted with EtOAc and washedwith water, brine and sat NaHCO₃. The organic layer was dried (MgSO₄),filtered and concentrated to afford a dark orange oil. The crudematerial was purified by silica column chromatography eluting with50-80% EtOAc/hexanes to afford (R)-ethyl5-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate(0.72 g, 56.2% yield) as a yellow foamy solid. MS (apci) m/z=386.0(M+H).

Step D: Preparation of(R)-5-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylicacid

To a suspension of (R)-ethyl5-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate(0.72 g, 1.868 mmol) in MeOH (9.34 mL) was added LiOH (1 N, 3.74 mL,3.74 mmol), and the reaction mixture was heated at 70° C. for 15 hours.After cooling, the reaction mixture was concentrated and the resultingresidue diluted in water. After acidifying with citric acid, the aqueouslayer was extracted with DCM. The combined organics were dried (MgSO₄),filtered and concentrated to afforded(R)-5-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylicacid (0.67 g, 100% yield) as a yellow solid. MS (apci) m/z=357.9 (M+H).

Preparation C

(R)-4-((tert-butyldimethylsilyl)oxy)-2-(5-fluoro-2-methoxyphenyl)pyrrolidine2,2,2-trifluoroacetate

Steps A-D followed the procedure reported by H. Imamura, et al. inTetrahedron, 2000, 56, 7705.

Step A: Preparation of(R)-4-(tert-butyldimethylsilyloxy)pyrrolidin-2-one

To a suspension of (R)-4-hydroxypyrrolidin-2-one (purchased from AstaTech or Aldrich) (5.030 g, 48.26 mmol) in DMF (24 mL) at 0° C. was addedTBDMS-Cl (7.637 g, 50.67 mmol) followed by imidazole (4.978 g, 72.39mmol). The resulting mixture was warmed to ambient temperature andstirred for 1 hour, then poured into 100 mL of water with stirring. Theresulting suspension was filtered and the solids were washed with waterand dried under reduced pressure to afford(R)-4-(tert-butyldimethylsilyloxy)pyrrolidin-2-one (10.14 g, 97.56%yield) that was used directly without further purification.

Step B: Preparation of (R)-tert-butyl4-(tert-butyldimethylsilyloxy)-2-oxopyrrolidine-1-carboxylate

To a solution of (R)-4-(tert-butyldimethylsilyloxy)pyrrolidin-2-one(10.14 g, 47.08 mmol) in MeCN (16 mL) at 0° C. was added sequentiallyDMAP (3.221 g, 26.37 mmol), TEA (3.957 mL, 28.25 mmol), and Boc₂O (11.49g, 52.65 mmol). The resulting mixture was warmed to ambient temperatureand stirred for 48 hours. The reaction mixture was poured into water andextracted with EtOAc (100 mL). The organic layer was successively washedwith 1 N aqueous HCl (2×50 mL), 1 N aqueous NaOH (50 mL), and brine. Theorganic layer was dried over MgSO₄, filtered, and concentrated in vacuoto afford (R)-tert-butyl4-(tert-butyldimethylsilyloxy)-2-oxopyrrolidine-1-carboxylate (13.62 g,91.69% yield). ¹H NMR (CDCl₃) δ 4.39 (m, 1H), 3.87 (m, 1H), 3.62 (m,1H), 2.71 (m, 1H), 2.46 (m, 1H), 1.53 (s, 9H), 0.88 (s, 9H), 0.08 (d,6H).

Step C: Preparation of (R)-tert-butyl2-(tert-butyldimethylsilyloxy)-4-(5-fluoro-2-methoxyphenyl)-4-hydroxybutylcarbamate

To a solution of (R)-tert-butyl4-(tert-butyldimethylsilyloxy)-2-oxopyrrolidine-1-carboxylate (6.00 g,19.0 mmol) in THF (36 mL) at 0° C. was added a 0.5 M solution of(5-fluoro-2-methoxyphenyl)magnesium bromide in THF (50.0 mL, 25.0 mmol).The resulting mixture was stirred at 0° C. for 30 minutes, then treatedwith MeOH (60 mL) and NaBH₄ (0.966 g, 25.2 mmol). After stirring at 0°C. for an additional 30 minutes, the reaction mixture was poured intosaturated aqueous NH₄Cl (40 mL) and extracted with EtOAc (2×50 mL). Thecombined organic layers were washed with brine, dried over MgSO₄,filtered, and concentrated under reduced pressure to give the crudematerial which was purified by silica column chromatography, elutingwith 0-2% MeOH/DCM to afford (R)-tert-butyl2-(tert-butyldimethylsilyloxy)-4-(5-fluoro-2-methoxyphenyl)-4-hydroxybutylcarbamate(which was assumed to be a mixture of the syn and anti isomers), (4.81g, 57.0% yield). ¹H NMR (CDCl₃) δ 7.20 (m, 1H), 6.90 (m, 1H), 6.77 (m,1H), 5.12 (m, 1H), 4.10 (m, 1H), 3.82 (m, 3H), 3.29 (m, 2H), 1.71-1.93(m, 2H), 1.45 (s, 9H), 0.93 (d, 9H), 0.11-0.14 (m, 6H).

Step D: Preparation of (R)-tert-butyl4-(tert-butyldimethylsilyloxy)-2-(5-fluoro-2-methoxyphenyl)pyrrolidine-1-carboxylate

To a solution of (R)-tert-butyl2-(tert-butyldimethylsilyloxy)-4-(5-fluoro-2-methoxyphenyl)-4-hydroxybutylcarbamate(4.810 g, 10.84 mmol) in CH₂Cl₂ (108 mL) at −60° C. was added TEA (4.534mL, 32.53 mmol) followed by methanesulfonyl chloride (0.9231 mL, 11.93mmol). The resulting mixture was slowly warmed to −5° C. and poured intoa mixture of ice and saturated aqueous NaHCO₃ (50 rnL). The organiclayer was separated and the aqueous layer was extracted with CH₂Cl₂(2×50 mL). The combined organic layers were dried over MgSO₄, filtered,and concentrated under reduced pressure to give the crude material thatwas purified by silica column chromatography, eluting with 2-10%MeOH/DCM to afford (R)-tert-butyl4-(tert-butyldimethylsilyloxy)-2-(5-fluoro-2-methoxyphenyl)pyrrolidine-1-carboxylate(assumed to be a mixture of cis and trans isomers; 2.648 g, 57.38%yield). LC/MS (ES+APCI) m/z=326.1 (M+H-Boc).

Step E: Preparation of(R)-4-(tert-butyldimethylsilyloxy)-2-(5-fluoro-2-methoxyphenyl)pyrrolidine2,2,2-trifluoroacetate

To a solution of (R)-tert-butyl4-(tert-butyldimethylsilyloxy)-2-(5-fluoro-2-methoxyphenyl)pyrrolidine-1-carboxylate(2.648 g, 6.222 mmol) in CH₂Cl₂ (26 mL) at 0° C. was added TFA (9.3 mL).The resulting mixture was warmed to ambient temperature and stirred for2 hours. The reaction mixture was concentrated under reduced pressure togive the crude material that was azeotroped with toluene-CH₂Cl₂ (2×) anddried under reduced pressure to provide(R)-4-(tert-butyldimethylsilyloxy)-2-(5-fluoro-2-methoxyphenyl)pyrrolidine2,2,2-trifluoroacetate (assumed to be a mixture of cis and transisomers; 2.92 g, 106.8% yield), which was used directly without furtherpurification. LC/MS (ES+APCI) m/z=326.3 (M+H).

Preparation D

(R)-5-(2-(5-fluoro-2-hydroxyphenyl)-4-hydroxypyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylicacid Step A: Preparation of (R)-ethyl5-(4-(tert-butyldimethylsilyloxy)-2-(5-fluoro-2-methoxyphenyl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate

To a suspension of ethyl5-hydroxypyrazolo[1,5-a]pyrimidine-3-carboxylate (0.100 g, 0.483 mmol)and BOP reagent (0.320 g, 0.724 mmol) in DMF (1 mL) at 0° C. was added asolution of(R)-4-(tert-butyldimethylsilyloxy)-2-(5-fluoro-2-methoxyphenyl)pyrrolidine2,2,2-trifluoroacetate (Preparation C; 0.167 g, 0.483 mmol) in CH₂Cl₂ (1mL) and N,N-diisopropylethylamine (0.420 mL, 2.41 mmol) sequentially.The resulting mixture was warmed to ambient temperature and stirred for18 hours. The reaction mixture was diluted with EtOAc (10 mL), andwashed with saturated aqueous NaHCO₃ and brine. The brine phase wasback-extracted with EtOAc (3×). The combined organic layers were driedover MgSO₄, filtered, and concentrated under reduced pressure to givethe crude material that was purified by silica column chromatography,eluting with 0-50% EtOAc/Hexanes to afford (R)-ethyl5-(4-(tert-butyldimethylsilyloxy)-2-(5-fluoro-2-methoxyphenyl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate(as a mixture of cis and trans isomers) (0.0487 g, 19.6% yield). LC/MS(ES+APCI) m/z=515.2 (M+H).

Step B: Preparation of (R)-ethyl5-(2-(5-fluoro-2-methoxyphenyl)-4-hydroxypyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate

To a solution of (R)-ethyl5-(4-(tert-butyldimethylsilyloxy)-2-(5-fluoro-2-methoxyphenyl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate(as a mixture of the cis and trans isomers) (0.0487 g, 0.0946 mmol) inTHF (1 mL) at 0° C. was added 1 M TBAF in THF (0.104 mL, 0.104 mmol).The reaction mixture was warmed to ambient temperature and stirred for2.5 hours. The reaction mixture was diluted with EtOAc (10 mL), washedwith brine, dried over MgSO₄, filtered, and concentrated under reducedpressure to afford the crude (R)-ethyl5-(2-(5-fluoro-2-methoxyphenyl)-4-hydroxypyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate(as a mixture of cis and trans isomers; 37.9 mg, 100% yield). LC/MS(ES+APCI) m/z=401.1 (M+H).

Step C: Preparation of (R)-ethyl5-(2-(5-fluoro-2-hydroxyphenyl)-4-hydroxypyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate

To a solution of (R)-ethyl5-(2-(5-fluoro-2-methoxyphenyl)-4-hydroxypyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate(as a mixture of cis and trans isomers; 0.0379 g, 0.0947 mmol) in CH₂Cl₂(1 mL) at 0° C. was added 1 M BBr₃ in CH₂Cl₇ (0.473 ml, 0.473 mmol). Theresulting mixture was warmed to ambient temperature for 25 hours, thendiluted with CH₂Cl₂ (10 mL) and poured into a mixture of ice andsaturated aqueous NaHCO₃ (15 mL). The organic layer was separated andthe aqueous layer acidified with 1N aqueous HCl until pH=5-6. Theaqueous layer was extracted with CH₂Cl₂ (3×) and the combined organiclayers were dried over MgSO₄, filtered, and concentrated under reducedpressure to give a mixture of(R)-5-(2-(5-fluoro-2-hydroxyphenyl)-4-hydroxypyrrolidin-1-yl)pyrazolopyrimidine-3-carboxylic acid (as a mixture of cis and trans isomers) and(R)-ethyl5-(2-(5-fluoro-2-hydroxyphenyl)-4-hydroxypyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate(as a mixture of cis and trans isomers). The mixture was dissolved inMeOH-THF (0.25 mL/0.75 mL) and treated with 1 N aqueous LiOH (0.474 mL,0.474 mmol). The resulting mixture was heated at 50° C. for 1 hour, thencooled to ambient temperature and acidified to pH 3 to 4 with 1 Naqueous HCl. The mixture was extracted with EtOAc (3×15 mL) and thecombined organic layers were dried over MgSO₄, filtered, andconcentrated under reduced pressure to give crude(R)-5-(2-(5-fluoro-2-hydroxyphenyl)-4-hydroxypyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylicacid (as a mixture of cis and trans isomers; 33.9 mg, 100% yield). LC/MS(ES+APCI) m/z=357.1 (M−H).

Example 1

(6R)-9-fluoro-2,11,15,19,20,23-hexaazapentacyclo[15.5.2.1^(7,11).0^(2,6).0^(20,24)]pentacosa-1(23),7,9,17(24),18,21-hexaene-16,25-dioneStep A: Preparation of (R)-ethyl5-(2-(5-fluoro-2-oxo-1,2-dihydropyridin-3-1-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate

To a mixture of (R)-ethyl5-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate(Preparation B, Step C; 0.92 g, 2.39 mmol) and Acetic acid (5.73 g, 95.5mmol) was added HBr (4.4 mL, 23.9 mmol, 33% in acetic acid). Thereaction mixture was heated at 90° C. for 2 hours. After cooling, thereaction mixture was treated with EtOAc, washed with water, saturatedNaHCO₃ and brine, then dried (MgSO₄), filtered, and concentrated. Thecrude material was purified by silica column chromatography, elutingwith 3% MeOH/DCM to yield the desired product (0.605 g, 68% yield). MS(apci) m/z=372.0 (M+H).

Step B: Preparation of (R)-ethyl5-(2-(1-(3-(1,3-dioxoisoindolin-2-yl)propyl)-5-fluoro-2-oxo-1,2-dihydropyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate

To a DMF (5 mL) suspension of (R)-ethyl5-(2-(5-fluoro-2-oxo-1,2-dihydropyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate(0.20 g, 0.54 mmol) was added LiH (6.8 mg, 0.81 mmol) at 0° C., followedfirst by 20-minute stirring, then addition of a DMF (1 mL) solution of2-(3-bromopropyl)isoindoline-1,3-dione (0.29 g, 1.1 mmol). The reactionwas warmed to ambient temperature and stirred for 17 hours. Aftercooling to 0° C. the reaction was quenched with ice-water (30 mL) andthe aqueous was extracted with EtOAc (3×50 mL). The combined organiclayers were backwashed with water and brine, dried (MgSO₄), filtered,and concentrated. The crude material was purified by silica columnchromatography, eluting with 2% MeOH/DCM to yield the desired product(0.2 g, 66% yield). MS (apci) m/z=559.0 (M+H).

Step C: Preparation of (R)-ethyl5-(2-(1-(3-aminopropyl)-5-fluoro-2-oxo-1,2-dihydropyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate

To a solution of (R)-ethyl5-(2-(1-(3-(1,3-dioxoisoindolin-2-yl)propyl)-5-fluoro-2-oxo-1,2-dihydropyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate(0.20 g, 0.36 mmol) in 1:1 MeOH/THF (12 mL) was added hydrazine-H₂O(0.18 g, 3.6 mmol). The reaction mixture was heated at 50° C. for 24hours. After cooling, the reaction mixture was poured into water andextracted with DCM (3×20 mL). The combined organics were dried (MgSO₄),filtered, and concentrated to afford the desired product (0.11 g, 72%yield). MS (apci) m/z=429.0 (M+H).

Step D: Preparation of(R)-5-(2-(1-(3-aminopropyl)-5-fluoro-2-oxo-1,2-dihydropyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylicacid

To a solution of (R)-ethyl5-(2-(1-(3-aminopropyl)-5-fluoro-2-oxo-1,2-dihydropyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate(0.11 g, 0.26 mmol) in 3:1 THF/MeOH (8 mL) was added LiOH (1 N, 1.5 mL,1.5 mmol), and the reaction mixture was heated at 70° C. for 20 hours.After cooling, the reaction mixture was treated with MeOH, acidifiedwith 1N HCl (1.5 mL), and concentrated to afford the desired product(0.1 g, 100% yield). MS (apci) m/z=401.1 (M+H).

Step E: Preparation of(6R)-9-fluoro-2,11,15,19,20,23-hexaazapentacyclo[15.5.2.1^(7,11).0^(2,6).0^(20,24)]pentacosa-1(23),7,9,17(24),18,21-hexaene-16,25-dione

To a solution of(R)-5-(2-(1-(3-aminopropyl)-5-fluoro-2-oxo-1,2-dihydropyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylicacid (95 mg, 0.24 mmol) in 1:2 DMF/DCM (9 mL) was added EDCI (0.14 g,0.71 mmol) followed by HOBT (96 mg, 0.71 mmol) at ambient temperature.After stirring for 10 minutes, TEA (0.099 mL, 0.71 mmol) was added tothe reaction mixture and stirred for 6 hours. The reaction mixture wastreated with EtOAc, washed with saturated NH₄Cl, saturated NaHCO₃, andbrine, then dried (MgSO₄), filtered, concentrated. The crude materialwas purified by silica column chromatography, eluting with 4% MeOH/DCMto yield the title product (35 mg, 39% yield). MS (apci) m/z=383.2(M+H).

Example 2

(6R)-12-oxa-2,16,20,21,24,26-hexaazapentacyclo[16.5.2.1^(7,11).0^(2,6).0^(21,25)]hexacosa-1(24),7(26),8,10,18(25),19,22-heptaen-17-oneStep A: Preparation of (R)-2-methoxy-6-(pyrrolidin-2-yl)pyridine

Prepared according to the method described in Preparation A,substituting 3-bromo-5-fluoro-2-methoxypyridine with2-bromo-6-methoxypyridine in Step A. MS (apci) m/z=179.1 (M+H).

Step B: Preparation of (R)-ethyl5-(2-(6-methoxypyridin-2-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate

Prepared by to the same method as described in Preparation B, Step C,substituting (R)-5-fluoro-2-methoxy-3-(pyrrolidin-2-yl)pyridine with(R)-2-methoxy-6-(pyrrolidin-2-yl)pyridine. MS (apci) m/z=368.0 (M+H).

Step C: Preparation of (R)-ethyl5-(2-(6-oxo-1,6-dihydropyridin-2-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate

To a mixture of (R)-ethyl5-(2-(6-methoxypyridin-2-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate(0.46 g, 1.25 mmol) and acetic acid (3.0 g, 50 mmol) was added HBr (3.1g, 12.5 mmol, 33% in acetic acid). The reaction mixture was heated at90° C. for 2 hours. After cooling, the reaction was diluted with EtOAc,washed with water, saturated NaHCO₃, and brine, then dried (MgSO₄),filtered, and concentrated. The crude material was purified by silicacolumn chromatography, eluting with 4% MeOH/DCM to yield the desiredproduct (0.3 g, 67% yield). MS (apci) m/z=354.1 (M+H).

Step D: Preparation of (R)-ethyl5-(2-(6-(3-(1,3-dioxoisoindolin-2-yl)propoxy)pyridin-2-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate

To a suspension of (R)-ethyl5-(2-(6-oxo-1,6-dihydropyridin-2-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate(0.091 g, 0.26 mmol) in DMF (2 mL) was added LiH (3.2 mg, 0.39 mmol) at0° C. After stirring for 20 minutes, a solution of2-(3-bromopropyl)isoindoline-1,3-dione (0.14 g, 0.52 mmol) in DMF (1 mL)was added, and the reaction was warmed up to ambient temperature andstirred for 17 hours. After cooling to 0° C., the reaction was quenchedwith ice-water (30 mL) and extracted with EtOAc (3×50 mL). The combinedorganic layers were washed with water and brine, dried (MgSO₄),filtered, and concentrated. The crude material was purified by silicacolumn chromatography, eluting with 1.5% MeOH/DCM to yield the desiredproduct (0.117 g, 84% yield). MS (apci) m/z=541.1 (M+H).

Step E: Preparation of (R)-ethyl5-(2-(6-(3-aminopropoxy)pyridin-2-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate

To a solution of (R)-ethyl5-(2-(6-(3-(1,3-dioxoisoindolin-2-yl)propoxy)pyridin-2-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate(0.11 g, 0.20 mmol) in 1:1 MeOH/THF (12 mL) was added hydrazine-H₂O(0.10 g, 2.0 mmol). The reaction mixture was heated at 50° C. for 24hours. After cooling, the reaction mixture was poured into water thenextracted with DCM (3×20 mL). The combined organics were dried (MgSO₄),filtered, and concentrated to afford the desired product (70 mg, 84%yield). MS (apci) m/z=441.1 (M+H).

Step F: Preparation of(R)-5-(2-(6-(3-aminopropoxy)pyridin-2-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylicacid

To a solution of (R)-ethyl5-(2-(6-(3-aminopropoxy)pyridin-2-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate(70 mg, 0.17 mmol) in 3:1 THF/MeOH (8 mL) was added LiOH (1 N, 1.5 mL,1.5 mmol) and the reaction mixture was heated at 70° C. for 20 hours.After cooling, the reaction mixture was diluted with MeOH, acidifiedwith 1 N HCl (1.5 mL), and concentrated to afford the desired product(65 mg, 100% yield). MS (apci) m/z=383.1 (M+H).

Step G: Preparation of(6R)-12-oxa-2,16,20,21,24,26-hexaazapentacyclo-[16.5.2.1^(7,11).0^(2,6).0^(21,25)]hexacosa-1(24),7(26),8,10,18(25),19,22-heptaen-17-one

To a solution of(R)-5-(2-(6-(3-aminopropoxy)pyridin-2-yl)pyrrolidin-1-yl)pyrazolopyrimidine-3-carboxylic acid (70 mg, 0.18 mmol) in 1:2 DMF/DCM (9 mL)was added EDCI (110 mg, 0.55 mmol) followed by HOBT (74 mg, 0.55 mmol)at ambient temperature. After stirring for 10 minutes, TEA (0.077 mL,0.55 mmol) was added to the reaction mixture and stirred for 6 hours.The reaction mixture was diluted with EtOAc, washed with saturatedNH₄Cl, saturated NaHCO₃, and brine, then dried (MgSO₄), filtered, andconcentrated. The crude material was purified by silica columnchromatography, eluting with 2% MeOH/DCM to yield the title product (30mg, 45% yield). MS (apci) m/z=365.2 (M+H).

Example 3

(6R)-9-fluoro-13-oxa-2,11,17,21,22,25-hexaazapentacyclo[17.5.2.0^(2,6).0^(7,12).0^(22,26)]hexacosa-1(25),7,9,11,19(26),20,23-heptaen-18-oneStep A: Preparation of(R)-5-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)-N-(3-hydroxypropyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

To a DMF (2 mL) suspension of(R)-5-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylicacid (Preparation B, 250 mg, 0.700 mmol) and HATU (319 mg, 0.840 mmol)cooled to 0° C. was added 3-aminopropan-1-ol (0.0642 mL, 0.840 mmol)drop-wise, resulting in a clear yellowish solution. After dropwiseaddition of DIEA (0.366 mL, 2.10 mmol), ice bath was removed andreaction was stirred at ambient temperature for 1 hour. The reaction wasdirectly purified on reverse phase column chromatography (Biotage SP4system, C-18 25+M column, 0 to 54% Acetonitrile/water), to provide theproduct as white solid (200 mg, 69% yield). MS (apci) m/z=415.1 (M+H).

Step B: Preparation of(R)—N-(3-chloropropyl)-5-(2-(5-fluoro-2-hydroxypyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

A mixture of(R)-5-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)-N-(3-hydroxypropyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide(20 mg, 0.0483 mmol) in HCl (4 N dioxane, 1.2 mL, 4.83 mmol) was heatedat 85° C. overnight. The reaction mixture was concentrated, trituratedwith ether, and filtered, to provide the crude product as a beige solid,which was directly used in the next step without further purification(22 mg, 106% yield). MS (apci) m/z=419.1 (M+H).

Step C: Preparation of(6R)-9-fluoro-13-oxa-2,11,17,21,22,25-hexaazapentacyclo[17.5.2.0^(2,6).0^(7,12).0^(22,26)]hexacosa-1(25),7,9,11,19(26),20,23-heptaen-18-one

A DMF (1 mL) suspension of(R)—N-(3-chloropropyl)-5-(2-(5-fluoro-2-hydroxypyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide(5 mg, 0.012 mmol) and Cs₂CO₁ (4 mg, 0.06 mmol) was heated at 85° C.overnight. The reaction mixture was filtered through a GF/F paper anddirectly purified on reverse phase column chromatography (Biotage SP4system C-18 12+M column, 5 to 60% acetonitrile/water), to provide thetitle product as white solid (2 mg, 44% yield). MS (apci) m/z=383.3(M+H).

Example 4

(6R)-9-fluoro-15-hydroxy-13-oxa-2,11,17,21,22,25-hexaazapentacyclo[17.5.2.0^(2,6).0^(7,12).0^(22,26)]hexacosa-1(25),7,9,11,19(26),20,23-heptaen-18-oneStep A: Preparation ofN-(2,3-dihydroxypropyl)-5-((R)-2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

A mixture of(R)-5-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)pyrazolopyrimidine-3-carboxylic acid (Preparation B, 250 mg, 0.700 mmol) andHATU (319 mg, 0.840 mmol) in 1:1 DMF/DMSO (2 mL) was cooled to 0° C.,followed first by drop-wise addition of 3-aminopropane-1,2-diol (76.5mg, 0.840 mmol) and then addition of DIEA (366 μL, 2.10 mmol). Thereaction was warmed up to ambient temperature, stirred for 20 minutes,and then directly purified on reverse phase column chromatography(Biotage SP4 system C-18 25+M cartridge, 5 to 50% acetonitrile/water),to provide the product as a white solid (295 mg, 98% yield). MS(apci)=431.1 (M+H).

Step B: Preparation ofN-(3-chloro-2-hydroxypropyl)-5-((R)-2-(5-fluoro-2-oxo-1,2-dihydropyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

A mixture ofN-(2,3-dihydroxypropyl)-5-((R)-2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide(100 mg, 0.232 mmol) and HCl (4 N, dioxane, 5.8 mL) was sealed in apressure tube and heated at 85° C. overnight. After the clear solutionwas decanted, the crude product was obtained as a brownish oily residue,which was vacuum-dried and used directly in the next step withoutfurther purification. MS (apci) m/z=435.0 (M+H).

Step C: Preparation of(6R)-9-fluoro-15-hydroxy-13-oxa-2,11,17,21,22,25-hexaazapentacyclo[17.5.2.0^(2,6).0^(7,12).0^(22,26)]hexacosa-1(25),7,9,11,19(26),20,23-heptaen-18-one

A suspension ofN-(3-chloro-2-hydroxypropyl)-5-((R)-2-(5-fluoro-2-oxo-1,2-dihydropyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide(100 mg, 0.23 mmol) and Cs₂CO₃ (375 mg, 1.15 mmol) in DMF (3 mL) washeated at 85° C. for 2 hours. The reaction mixture was filtered througha GF/F paper and directly purified on reverse phase columnchromatography (Biotage SP4 system C-18 25+M column, 5 to 50%acetonitrile/water), to provide the title product as a white solid. MS(apci) m/z=399.2.

Example 5

(6R,13S)-9-fluoro-13-hydroxy-2,11,15,19,20,23-hexaazapentacyclo-[15.5.2.1^(7,11).0^(2,6).0^(20,24)]pentacosa-1(23),7,9,17(24),18,21-hexaene-16,25-dioneStep A: Preparation ofN—((S)-3-chloro-2-hydroxypropyl)-5-((R)-2-(5-fluoro-2-oxo-1,2-dihydropyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamidehydrochloride

Prepared according to the method described in Example 3, Steps A-B,substituting 3-aminopropan-1-ol in Step A with(S)-3-aminopropane-1,2-diol. MS (apci) m/z=435.0 (M+H).

Step B: Preparation of(6R,13S)-9-fluoro-13-hydroxy-2,11,15,19,20,23-hexaazapentacyclo[15.5.2.1^(7,11).0^(2,6).0^(20,24)]pentacosa-1(23),7,9,17(24),18,21-hexaene-16,25-dione

A suspension ofN—((S)-3-chloro-2-hydroxypropyl)-5-4R)-2-(5-fluoro-2-oxo-1,2-dihydropyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamidehydrochloride (40 mg, 0.085 mmol) and Cs₂CO₃ (138 mg, 0.42 mmol) in DMF(0.8 mL) was heated at 85° C. for 2 hours. The reaction mixture wasfiltered through GF/F paper and directly purified on reverse phasecolumn chromatography (Biotage SP4 system C-18 12+M column, 0 to 40%acetonitrile/water), to provide the title product as a white solid (4mg, 12% yield). MS (apci) m/z=399.2 (M+H).

Example 6

(6R)-9-fluoro-15-hydroxy-13-oxa-2,11,17,21,22,25-hexaazapentacyclo-[17.5.2.0^(2,6).0^(7,12).0^(22,26)]hexacosa-1(25),7,9,11,19(26),20,23-heptaen-18-one

Prepared according to the method described in Example 5 and isolated asa by-produce in Step B. The enantiomeric integrity of the chiral centerwhere the HO group resides was found to have unexpectedly eroded (R/Sratio was about 10:7) in the isolated final product(6R)-9-fluoro-15-hydroxy-13-oxa-2,11,17,21,22,25-hexaazapentacyclo-[17.5.2.0^(2,6).0^(7,12).0^(22,26)]hexacosa-1(25),7,9,11,19(26),20,23-heptaen-18-one,which was obtained as a white solid (5 mg, 15% yield) by reverse phasecolumn chromatography (Biotage SP4 system C-18 12+M column, 0 to 50%acetonitrile/water). MS (apci) m/z=399.2 (M+H).

Example 7

(6R,15R)-9-fluoro-15-hydroxy-13-oxa-2,11,17,21,22,25-hexaazapentacyclo-[17.5.2.0^(2,6).0^(7,12).0^(22,26)]hexacosa-1(25),7,9,11,19(26),20,23-heptaen-18-oneStep A: Preparation ofN—((R)-3-chloro-2-hydroxypropyl)-5-((R)-2-(5-fluoro-2-oxo-1,2-dihydropyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

Prepared according to the method described in Example 3, Steps A-B,substituting 3-aminopropan-1-ol in Step A with(R)-3-aminopropane-1,2-diol. MS (apci) m/z=435.0 (M+H).

Step B: Preparation of(6R,15R)-9-fluoro-15-hydroxy-13-oxa-2,11,17,21,22,25-hexaazapentacyclo[17.5.2.0^(2,6).0^(7,12).0^(22,26)]hexacosa-1(25),7,9,11,19(26),20,23-heptaen-18-one

A suspension ofN—((R)-3-chloro-2-hydroxypropyl)-5-((R)-2-(5-fluoro-2-oxo-1,2-dihydropyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide(30 mg, 0.069 mmol) and Cs₂CO₃ (112 mg, 0.34 mmol) in DMF (0.7 mL) washeated at 85° C. for 1 hour. The reaction mixture was filtered through aGF/F paper and directly purified on reverse phase column chromatography(Biotage SP4 system C-18 12+M column, 0 to 50% acetonitrile/water), toprovide the title product as a white solid (10 mg, 36% yield). UnlikeExample 6, no erosion of the enantiomeric integrity of the chiral centerwhere HO group resides was observed for this final product. MS (apci)m/z=399.2 (M+H).

Example 8

(6R,13R)-9-fluoro-13-hydroxy-2,11,15,19,20,23-hexaazapentacyclo-[15.5.2.1^(7,11).0^(2,6).0^(20,24)]pentacosa-1(23),7,9,17(24),18,21-hexaene-16,25-dione

Obtained as a by-product of Example 7, Step B and isolated as a whitesolid (1.2 mg, 4% yield) by reverse phase column chromatography (BiotageSP4 system C-18 12+M column, 0 to 44% acetonitrile/water) of the crudematerial of Example 7, Step B. MS (apci) m/z=399.2 (M+H).

Example 9

(6R)-9-fluoro-13-oxa-2,11,16,20,21,24-hexaazapentacyclo[16.5.2.0^(2,6).0^(7,12).0^(21,25)]pentacosa-1(24),7,9,11,18(25),19,22-heptaen-17-one Step A: Preparation of(R)-5-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)-N-(2-hydroxyethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

To a DMF (1 mL) suspension of(R)-5-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylicacid (Preparation B, 100 mg, 0.28 mmol) and HATU (128 mg, 0.336 mmol)was added DIEA (0.146 mL, 0.840 mmol) at ambient temperature, followedby a solution of 2-aminoethanol (20.5 mg, 0.336 mmol) in minimal amountof DMF dropwise at 0° C. The reaction was warmed up to ambienttemperature and stirred for 30 minutes, then directly purified byreverse-phase column chromatography (0 to 70% acetonitrile/water) toyield the product as white solid (95 mg, 85% yield). MS (apci pos)m/z=401.1 (M+H).

Step B: Preparation of(R)—N-(2-chloroethyl)-5-(2-(5-fluoro-2-oxo-1,2-dihydropyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

To(R)-5-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)-N-(2-hydroxyethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide(77 mg, 0.192 mmol) in a pressure reaction tube was charged hydrogenchloride (4 N dioxane, 4.8 mL, 19 2 mmol) and the resulting whitesuspension was heated at 85° C. overnight. After cooling to ambienttemperature, the reaction mixture was decanted to yield the crudeproduct as brownish oily residue, which was dried in vacuo and directlyused in the next step without further purification. MS (apci) m/z=405.0(M+H).

Step C: Preparation of(6R)-9-fluoro-13-oxa-2,11,16,20,21,24-hexaazapentacyclo[16.5.2.0^(2,6).0^(7,12).0^(21,25)]pentacosa-1(24),7,9,11,18(25),19,22-heptaen-17-one

A suspension of(R)—N-(2-chloroethyl)-5-(2-(5-fluoro-2-oxo-1,2-dihydropyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide(78 mg, 0.19 mmol) and Cs₂CO₃ (314 mg, 0.96 mmol) in DMF (5 mL) washeated at 85° C. for 30 minutes. After filtering through a GF/F paperthe reaction was diluted with water (40 mL) and NH₄Cl (saturated, 5 mL),then extracted with EtOAc (3×40 mL). The combined organic extracts weredried (Na₂SO₄), filtered, and concentrated. The crude material waspurified by reverse phase column chromatography (Biotage SP4 system C-1812+M column, 0 to 73% acetonitrile/water), to provide the title productas a white solid (17 mg, 24% yield). MS (apci) m/z=369.2 (M+H).

Example 10

(6R)-9-fluoro-13-oxa-2,11,18,22,23,26-hexaazapentacyclo[18.5.2.0^(2,6).0^(7,12).0^(23,27)]heptacosa-1(26),7,9,11,20(27),21,24-heptaen-19-oneStep A: Preparation of(R)—N-(4-chlorobutyl)-5-(2-(5-fluoro-2-oxo-1,2-dihydropyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

Prepared according to the method described in Example 3, Steps A-B,substituting 3-aminopropan-1-ol in Step A with 4-aminobutan-1-ol. MS(apci) m/z=433.0 (M+H).

Step B: Preparation of(6R)-9-fluoro-13-oxa-2,11,18,22,23,26-hexaazapentacyclo-[18.5.2.0^(2,6).0^(7,12).0^(23,27)]heptacosa-1(26),7,9,11,20(27),21,24-heptaen-19-one

Prepared according to the method described in Example 3, substituting(R)—N-(3-chloropropyl)-5-(2-(5-fluoro-2-hydroxypyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamidewith(R)—N-(4-chlorobutyl)-5-(2-(5-fluoro-2-oxo-1,2-dihydropyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamidein Step C. The crude product was purified on reverse phase columnchromatography (Biotage SP4 system C-18 25+M column, 0 to 80%acetonitrile/water), to provide the title product as a white solid (32mg, 44%). MS (apci pos) m/z=397.2 (M+H).

Example 11

(6R)-9-fluoro-2,11,16,20,21,24-hexaazapentacyclo[16.5.2.1^(7,11).0^(2,6).0^(21,25)]hexacosa-1(24),7,9,18(25),19,22-hexaene-17,26-dione

Obtained as a by-product in Example 10, Step B and isolated a whitesolid (4 mg, 6%) upon purification of the crude material of Example 10,Step B by reverse phase column chromatography (Biotage SP4 system C-1825+M column, 0 to 50% acetonitrile/water). MS (apci) m/z=397.2 (M+H).

Example 12

(6R)-9-fluoro-2,11,13,16,20,21,24-heptaazapentacyclo[16.5.2.0^(2,6).0^(7,12).0^(21,25)]pentacosa-1(24),7,9,11,18(25),19,22-heptaen-17-oneStep A: Preparation of (R)-tert-butyl2-(2-chloro-5-fluoropyridin-3-yl)pyrrolidine-1-carboxylate

A solution of tert-butyl pyrroidine-1-carboxylate (1 mL 5.70 mmol) and(−)-sparteine (1.31 mL, 5.70 mmol) in anhydrous MTBE (30 mL) was firstcooled to −78° C. under nitrogen, followed by addition of sec-butyllithium (4.07 mL, 1.4M, 5.70 mmol) drop-wise over 15 minutes with asyringe, maintaining the temperature below −75° C. The pale yellowishsolution was stirred at −78° C. for 3 hours before being treated withzinc chloride (3.80 mL, 1.0 M, 3.80 mmol) drop-wise over 15 minutes,maintaining the temperature below −73° C. The mixture was stirred at−78° C. for 30 minutes, then placed into an ambient temperature waterbath and stirred for another hour. At this point a large amount of whiteprecipitate was present. The mixture was treated with3-bromo-2-chloro-5-fluoropyridine (1.00 g, 4.75 mmol) in MTBE (5 mL),followed by addition of palladium acetate (53 mg, 0.24 mmol) andtri-t-butylphosphine tetrafluoroborate (83 mg, 0.28 mmol). The mixturewas allowed to stir at ambient temperature overnight to reachcompletion. The mixture was treated with NH₄OH (1 mL), stirred for 30minutes and filtered through GET paper, washing with MTBE. The filtratewas washed with 10% citric acid (30 mL) and the aqueous layer wasback-washed with MTBE (2×30 mL). The combined organic phases were washedwith brine (20 mL), dried (MgSO₄), and concentrated to afford the crudeproduct as dark yellowish oil. This crude material was purified on asilica 50 g Biotage SNAP cartridge eluting with 10% EtOAc in hexanes toafford the desired product as colorless oil (0.5 g, 35% yield). MS(apci) m/z=201.1 (M+H-Boc).

Step B: Preparation of (R)-2-chloro-5-fluoro-3-(pyrrolidin-2-yl)pyridinedihydrochloride

To a dioxane (5 mL) solution of (R)-tert-butyl2-(2-chloro-5-fluoropyridin-3-yl)pyrrolidine-1-carboxylate (500 mg, 1.66mmol) was added HCl (4 N dioxane, 20 mL), followed by stirring atambient temperature overnight. The mixture was concentrated and treatedwith Et₂O, then vacuum-dried, to provide the product as a white solid(0.36 g, 80% yield). MS (apci) m/z=201.1 (M+H). The enantiomeric excess(ee %) of the product was determined to be >92% according to the methoddescribed in Preparation A.

Step C: Preparation of (R)-ethyl5-(2-(2-chloro-5-fluoropyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate

To a solution of ethyl 5-hydroxypyrazolo[1,5-a]pyrimidine-3-carboxylate(Preparation B, Step A, 275 mg, 1.33 mmol) in anhydrous DMF (5 mL) wasadded (Benzotriazol-1-yloxy)tris(dimethylamino)phosphoniumhexafluorophosphate (BOP) (646 mg, 1.46 mmol). The heterogeneous mixturewas stirred for 10 minutes before adding DIEA (1.16 mL, 6.6 mmol),followed by addition of(R)-2-chloro-5-fluoro-3-(pyrrolidin-2-yl)pyridine dihydrochloride (363mg, 1.33 mmol). The reaction was stirred at ambient temperatureovernight to reach completion. The mixture was partitioned between 10%citric acid (30 mL) and EtOAc (30 mL), and the aqueous layer wasextracted with EtOAc (2×20 mL). The combined organic phases were washedsuccessively with water (20 mL), saturated NaHCO₃ (20 mL), water (20 mL)and brine (3×20 mL), then dried (Na₂SO₄) and concentrated to afford thecrude product as an orange foam. The crude material was purified on a 25g Biotage SNAP silica cartridge eluting with 1% MeOH/DCM to afford thedesired product as cream-colored foam (0.35 g, 68% yield). MS (apci)m/z=390.0 (M+H).

Step D: Preparation of (R)-ethyl5-(2-(2-(2-(tert-butoxycarbonylamino)ethylamino)-5-fluoropyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate

A mixture of Pd₂dba₃ (7.05 mg, 0.00770 mmol), Cs₂CO₃ (125 mg, 0.385mmol), rac-Binap (19.2 mg, 0.0308 mmol), (R)-ethyl5-(2-(2-chloro-5-fluoropyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate(50 mg, 0.128 mmol), and tert-butyl 2-aminoethylcarbamate (24.7 mg,0.154 mmol) in degassed toluene (1 mL) was first purged with nitrogen,then sealed and subjected to microwave irradiation (120° C.) for 16hours. After cooled to ambient temperature, the reaction mixture wasdiluted with EtOAc (10 mL) and washed with water (2×5 mL). The organicwas dried (Na₂SO₄) and concentrated. The crude material was purified byreverse phase column chromatography (Biotage SP4 system C18 12+Mcartridge, 5 to 70% acetonitrile/water) to yield the desired product aswhite foamy solid (38 mg, 58% yield). MS (apci) m/z=514.1 (M+H).

Step E: Preparation of(R)-5-(2-(2-(2-(tert-butoxycarbonylamino)ethylamino)-5-fluoropyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylicacid

To a solution of (R)-ethyl5-(2-(2-(2-(tert-butoxycarbonylamino)ethylamino)-5-fluoropyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate(38 mg, 0.074 mmol) in THF/MeOH/water (2:2:1, 0.7 mL) was added LiOH—H₂O(9.3 mg, 0.22 mmol), followed by stirring at 50° C. for 18 hours. Afterremoval of solvent, the reaction residue was taken up in water (0.5 mL),and acidified with 1 N HCl (0.22 mL) to pH 3. The reaction mixture wasextracted with EtOAc (3×2 mL), dried (Na₂SO₄), filtered and concentratedto give the desired product, which was used in the next step directlywithout further purification, assuming quantitative conversion. MS(apci)=486.0 (M+H).

Step F: Preparation of(R)-5-(2-(2-(2-aminoethylamino)-5-fluoropyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylicacid hydrochloride

A solution of (R)-5-(2-(2-(2-(tert-butoxycarbonylamino)ethylamino)-5-fluoropyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylicacid (31 mg, 0.064 mmol) in HCl (4 N dioxane, 798 μL) and TFA (50% DCM,2 mL) was stirred at ambient temperature for 1 hour before it wasconcentrated and dried under high vacuum to yield to give the desiredproduct as off-white solid, which was used in the next step directlywithout further purification, assuming quantitative conversion. MS(apci) m/z=386.1 (M+H).

Step G: Preparation of(6R)-9-fluoro-2,11,13,16,20,21,24-heptaazapentacyclo-[16.5.2.0^(2,6).0^(7,12).0^(21,25)]pentacosa-1(24),7,9,11,18(25),19,22-heptaen-17-one

To a DMF (3 mL) solution of(R)-5-(2-(2-(2-aminoethylamino)-5-fluoropyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylicacid (25 mg, 0.065 mmol) was first added HATU (29 mg, 0.077 mmol),followed by five-minute stirring and then drop-wise addition of DTEA (56μL, 0.32 mmol). After stirring at ambient temperature overnight, thereaction was directly purified by reverse phase column chromatography(Biotage SP4 system C18 25+M cartridge, acetonitrile/water 5 to 45%), toyield the title product as off-white solid (7 mg, 30% yield). MS (apci)m/z=368.2 (M+H).

Example 13

(6R)-9-fluoro-2,11,13,17,21,22,25-heptaazapentacyclo[17.5.2.0^(2,6).0^(7,12).0^(22,26)]hexacosa-1(25),7,9,11,19(26),20,23-heptaen-18-oneStep A: Preparation of (R)-ethyl 5-(2-(2-(3-(tert-butoxycarbonylamino)propylamino)-5-fluoropyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate

Prepared according to the method described in Example 12, Steps D,substituting tert-butyl 2-aminoethylcarbamate with tert-butyl3-aminopropylcarbamate. MS (apci) m/z=528.1 (M+H).

Step B: Preparation of(6R)-9-fluoro-2,11,13,17,21,22,25-heptaazapentacyclo-[17.5.2.0^(2,6).0^(7,12).0^(22,26)]hexacosa-1(25),7,9,11,19(26),20,23-heptaen-18-one

Prepared according to the method described in Example 12, Steps E-G, inthree steps, from (R)-ethyl5-(2-(2-(3-(tert-butoxycarbonylamino)propylamino)-5-fluoropyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylateobtained above. The crude product was purified by reverse phase columnchromatography (Biotage SP4 system C-18 25+M cartridge, 5 to 50%acetonitrile/water), to provide the title product as white solid (6 mg,44% yield). MS (apci pos) m/z=382.2 (M+H).

Example 14

(6R)-9-fluoro-13,16-dioxa-2,11,20,21,24-pentaazapentacyclo[16.5.2.0^(2,6).0^(7,12).0^(21,25)]pentacosa-1(24),7,9,11,18(25),19,22-heptaen-17-oneStep A: Preparation of (R)-2-chloroethyl5-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate

To a DMF (1 mL) suspension of(R)-5-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylicacid (Preparation B, 0.1 g, 0.28 mmol) and HATU (0.128 g, 0.336 mmol)was added DIEA (0.146 ml, 0.840 mmol), followed by 2-chloroethanol(0.0270 g, 0.336 mmol). After stirring at ambient temperature for 30minutes, the reaction was directly purified by reverse phase columnchromatography (Biotage SP4 system C18 25+M, 5 to 65%acetonitrile/water) to obtain the intermediate(R)-3H-[1,2,3]triazolo[4,5-b]pyridin-3-yl5-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylateas white solid (94.7 mg, 71% yield). This isolated intermediate wasdissolved in excess chloroethanol (1 mL), followed by addition of dropsof DIEA at ambient temperature and stirred overnight to reachcompletion. The reaction was directly purified by reverse phase columnchromatography (Biotage SP4 system C18 25+M, acetonitrile/water 5 to 73)to obtain the titled product as white foamy solid (56 mg, 48% yield). MS(apci) m/z=419.9 (M+H).

Step B: Preparation of (R)-2-chloroethyl5-(2-(5-fluoro-2-oxo-1,2-dihydropyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate

A mixture of (R)-2-chloroethyl5-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate(56 mg, 0.13 mmol) in HCl (4 N dioxane, 2.5 mL, 10 mmol) was sealed in apressure reaction tube and heated at 100° C. for 45 minutes. Thereaction mixture was cooled and concentrated to yield the product asyellowish oil, which was used directly in the next step without furtherpurification, assuming quantitative yield. MS (apci) m/z=406.0 (M+H).

Step C: Preparation of(6R)-9-fluoro-13,16-dioxa-2,11,20,21,24-pentaazapentacyclo[16.5.2.0^(2,6).0^(7,12).0^(21,25)]-pentacosa-1(24),7,9,11,18(25),19,22-heptaen-17-one

A mixture of (R)-2-chloroethyl5-(2-(5-fluoro-2-oxo-1,2-dihydropyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate(54 mg, 0.133 mmol) and Cs₂CO₃ (217 mg, 0.665 mmol) in DMF (6 mL) washeated at 90° C. overnight. The reaction was filtered (GF/F paper) anddirectly purified by reverse phase column chromatography (Biotage SP4system C18 25+M, 5 to 60% acetonitrile/water) to yield a mixture ofdesired product and impurities. This mixture was treated with a secondcolumn chromatography on Biotage SNAP KP-Sil 10 g, eluting with 10%hexanes/EtOAc to give the pure title product as a white solid (11 mg,22% yield). MS (apci pos) m/z=370.2 (M+H).

Example 15

(6R)-9-fluoro-14-oxa-2,11,18,19,22-pentaazapentacyclo[14.5.2.1^(7,11).0^(2,6).0^(19,23)]tetracosa-1(22),7,9,16(23),17,20-hexaene-15,24-dione

Obtained as a by-product of Example 14, Step C, and isolated as a whitesolid (5 mg, 9% yield) by reverse phase column chromatography (BiotageSP4 system C-18 25+M column, 5 to 60% acetonitrile/water) of the crudematerial of Example 14, Step C. MS (apci) m/z=370.2 (M+H).

Example 16

(6R)-9-fluoro-13,16-dioxa-2,11,17,21,22,25-hexaazapentacyclo[17.5.2.0^(2,6).0^(7,12).0^(22,26)]hexacosa-1 (25),7,9,11,19(26),20,23-heptaen-18-one Step A:Preparation of(R)—N-(2-bromoethoxy)-5-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

To a mixture of(R)-5-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylicacid (Preparation B, 100 mg, 0.280 mmol) and HATU (128 mg, 0.336 mmol)in DMF (1 mL) was added DIEA (0.146 mL, 0.840 mmol), followed byO-(2-bromoethyl)hydroxylamine hydrobromide (74.2 mg, 0.336 mmol) in oneportion. After stirring at ambient temperature overnight, the reactionmixture was directly purified by reverse phase column chromatography(Biotage SP4 system C-18 25+M, 5 to 67% acetonitrile/water) to yield thedesired product as off-white solid (91 mg, 68% yield). MS (apci)m/z=479.0 (M+H).

Step B: Preparation of(R)—N-(2-chloroethoxy)-5-(2-(5-fluoro-2-oxo-1,2-dihydropyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

A mixture of(R)—N-(2-bromoethoxy)-5-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide(70 mg, 0.146 mmol) and HCl (4 N dioxane, 3.65 mL, 14.6 mmol) was sealedin a pressure tube and heated at 90° C. for 3 hours. The reactionmixture was then cooled, diluted with MeOH, concentrated, and dried onhigh vacuum to obtain the desired product which was used in the nextstep directly without further purification, assuming quantitativeconversion.

Step C: Preparation of(6R)-9-fluoro-13,16-dioxa-2,11,17,21,22,25-hexaazapentacyclo[17.5.2.0^(2,6).0^(7,12).0^(22,26)]hexacosa-1(25),7,9,11,19(26),20,23-heptaen-18-one

A mixture of(R)—N-(2-chloroethoxy)-5-(2-(5-fluoro-2-oxo-1,2-dihydropyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide(60 mg, 0.14 mmol) and Cs₂CO₃ (232 mg, 0.71 mmol) in DMF (1.4 mL) washeated at 90° C. for 20 minutes to reach completion. The reactionmixture was filtered (GF/F paper) and diluted with water (10 mL), thenextracted with EtOAc (3×10 mL). The organic layers were combined, washedwith brine and dried (Na₂SO₄). The crude material was purified onreverse phase column chromatography (Biotage SP4 system C18 12+M,acetonitrile/water 5 to 55%) to yield a mixture of the desired finalproduct and impurities. This mixture was again purified by preparativeTLC (10% MeOH/DCM) to yield the pure title product as white solid (1 mg,1% yield). MS (apci) m/z=385.1 (M+H).

Example 17

(6R,13R)-9,13-difluoro-2,11,15,19,20,23-hexaazapentacyclo[15.5.2.1^(7,11).0^(2,6).0^(20,24)]pentacosa-1(23),7,9,17(24),18,21-hexaene-16,25-dione

A solution of(6R,13S)-9-fluoro-13-hydroxy-2,11,15,19,20,23-hexaazapentacyclo-[15.5.2.1^(7,11).0^(2,6).0^(20,24)]pentacosa-1(23),7,9,17(24),18,21-hexaene-16,25-dione(Example 5; 10 mg, 0.0251 mmol) in a mixture solvent of DCM (0.3 mL) and3 drops of DMSO was treated with bis(2-methoxyethyl)amino-sulfurtrifluoride (7.87 μL, 0.0427 mmol) at 0° C., followed by addition of aDCM (0.1 mL) solution of ethanol (0.231 mg, 0.00502 mmol), and themixture was stirred at ambient temperature overnight. The reactionmixture was poured into saturated NaHCO₃ and extracted with DCM, thendried (Na₂SO₄), filtered, and concentrated. The crude material waspurified by reverse phase column chromatography (Biotage SP4 system C1812+M cartridge, acetonitrile/water 5 to 50%) to give the title productas beige solid (1.3 mg, 12% yield). MS (apci)=401.2 (M+H).

Example 18

(6R)-9-fluoro-17-methyl-13-oxa-2,11,17,21,22,25-hexaazapentacyclo[17.5.2.0^(2,6).0^(7,12).0^(22,26)]hexacosa-1(25),7,9,11,19(26),20,23-heptaen-18-oneStep A: Preparation of(R)—N-(3-chloropropyl)-5-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)-N-methylpyrazolopyrimidine-3-carboxamide

To a suspension of(R)-5-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylicacid (Preparation B, 200 mg, 0.56 mmol) and3-chloro-N-methylpropan-1-amine hydrochloride (177 mg, 1.23 mmol) in DMF(4 mL) was added N-methylmorpholine (0.25 mL, 2.30 mmol), followed byHATU (234 mg, 0.616 mmol). The reaction was stirred at ambienttemperature for 18 hours, then diluted with H₂O (10 mL), and extractedEtOAc (2×20 mL). The combined organic extracts were washed with brine(20 mL), dried (MgSO₄), filtered, and concentrated. The crude productwas purified by reverse-phase column chromatography, eluting with 5 to60% acetonitrile/water to yield the desired product as a white foamysolid (129 mg, 52% yield). MS (apci) m/z=447.0 (M+H).

Step B: Preparation of(R)—N-(3-chloropropyl)-5-(2-(5-fluoro-2-oxo-1,2-dihydropyridin-3-yl)pyrrolidin-1-yl)-N-methylpyrazolo[1,5-a]pyrimidine-3-carboxamide

A mixture of HCl (4 N dioxane, 4 mL, 16.0 mmol) and(R)—N-(3-chloropropyl)-5-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)-N-methylpyrazolo[1,5-a]pyrimidine-3-carboxamide(100 mg, 0.224 mmol) was sealed in a pressure tube and heated at 90° C.for 90 minutes. The reaction mixture was then diluted with acetonitrileand concentrated to yield the crude product, which was carried to thenext step without further purification (145 mg, 150% yield). MS (apci)m/z=433.0 (M+H).

Step C: Preparation of(6R)-9-fluoro-17-methyl-13-oxa-2,11,17,21,22,25-hexaazapentacyclo[17.5.2.0^(2,6).0^(7,12).0^(22,26)]hexacosa-1(25),7,9,11,19(26),20,23-heptaen-18-one

A mixture of(R)—N-(3-chloropropyl)-5-(2-(5-fluoro-2-oxo-1,2-dihydropyridin-3-yl)pyrrolidin-1-yl)-N-methylpyrazolo[1,5-a]pyrimidine-3-carboxamide(50 mg, 0.12 mmol) and Cs₂CO₃ (188 mg, 0.58 mmol) in DMF (12 mL) washeated at 90° C. for 15 minutes to reach completion. The reactionmixture was filtered, rinsed with DMF, and concentrated. The crudematerial was purified directly by reverse-phase column chromatography,eluting with 5 to 60% acetonitrile/water to yield the title product aspale yellow powder (17 mg, 36% yield). MS (apci) m/z=397.3 (M+H).

Example 19

(6R)-9,15,15-trifluoro-13-oxa-2,11,17,21,22,25-hexaazapentacyclo[17.5.2.0^(2,6).0^(7,12).0^(22,26)]hexacosa-1(25),7,9,11,19(26),20,23-heptaen-18-oneStep A: Preparation of (S)-1-amino-3-chloropropan-2-ol hydrochloride

To a solution of benzaldehyde (4.50 g, 42.4 mmol) in EtOH (12 mL) wasadded aqueous ammonia (4.01 g, 65.9 mmol) in several portions. Afterstirring for 10 minutes, (S)-2-(chloromethyl)oxirane (3.81 g, 41.2 mmol)was added and the reaction mixture was stirred for 2 hours at ambienttemperature. The reaction mixture was then heated at 35-40° C. with aheating mantle for 6 hours, followed by stirring at ambient temperaturefor 18 hours. The reaction was concentrated to 5 mL and toluene (5 mL)was added. The mixture was heated to 36° C. and a solution ofconcentrated HCl (6.09 g, 61.8 mmol) and water (5.9 mL) were addedslowly over 5 minutes to maintain an internal reaction temperature rangeof 36-41° C. The biphasic mixture was heated at 42-45° C. for 3 hours.The organic phase was separated and washed with water (10 mL). Theaqueous phases were combined and ethanol (10 mL) was added. The mixturewas concentrated to 10 mL, and ethanol (6×10 mL) was added,concentrating after each addition. After the last concentration step,the slurry was warmed to reflux, cooled to ambient temperature, and thenplaced at −20° C. for 18 hours. The product was collected by vacuumfiltration, washed with cold ethanol, and vacuum-dried, to provide theproduct as white crystalline solid (3.58 g, 60% yield). ¹H NMR (d⁶-DMSO)δ 8.14 (s, 3H), 5.91 (s, 1H), 3.93 (m, 1H), 3.59 (m, 2H), 2.89 (m, 1H),2.69 (m, 1H).

Step B: Preparation ofN—((S)-3-chloro-2-hydroxypropyl)-5-((R)-2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

Prepared according to the method described in Example 18, substituting(5)-1-amino-3-chloropropan-2-ol hydrochloride (98.1 mg, 0.672 mmol) for3-chloro-N-methylpropan-1-amine hydrochloride in Step A. MS (apci)m/z=448.9 (M+H).

Step C: Preparation of(R)—N-(3-chloro-2-oxopropyl)-5-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

To a solution ofN—((S)-3-chloro-2-hydroxypropyl)-5-((R)-2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide(180 mg, 0.401 mmol) in DCM (3 mL) was added Dess-Martin periodinane(204 mg, 0.481 mmol). The reaction was stirred at ambient temperaturefor 3 hours, then purified directly by reverse-phase columnchromatography, eluting with 5 to 60% acetonitrile/water to yield thedesired product as a white foamy solid (114 mg, 64% yield). MS (apci)m/z=447.0 (M+H).

Step D: Preparation of(R)—N-(3-chloro-2,2-difluoropropyl)-5-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

To a solution of(R)—N-(3-chloro-2-oxopropyl)-5-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide(114 mg, 0.255 mmol) in DCM (3 mL) was added Deoxofluor (0.103 mL, 0.561mmol), and the reaction mixture was stirred at ambient temperature for23 hours. The reaction was quenched with saturated NaHCO₃ (5 mL),diluted with DCM (5 mL), and stirred for 30 minutes. After phaseseparation, the aqueous phase was extracted with DCM (10 mL). Thecombined organic phases were concentrated and purified by reverse-phasecolumn chromatography, eluting with 5 to 60% acetonitrile/water to yieldthe desired product as white solid (59 mg, 49% yield). MS (apci)m/z=469.0 (M+H).

Step E: Preparation of(R)—N-(3-chloro-2,2-difluoropropyl)-5-(2-(5-fluoro-2-oxo-1,2-dihydropyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

Prepared according to the method described in Example 18, substituting(R)—N-(3-chloro-2,2-difluoropropyl)-5-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamidefor(R)—N-(3-chloropropyl)-5-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)-N-methylpyrazolo[1,5-a]pyrimidine-3-carboxamidein Step B. MS (apci) m/z=455.0 (M+H).

Step F: Preparation of(6R)-9,15,15-trifluoro-13-oxa-2,11,17,21,22,25-hexaazapentacyclo[17.5.2.0^(2,6).0^(7,12).0^(22,26)]hexacosa-1(25),7,9,11,19(26),20,23-heptaen-18-one

Prepared according to the same method as described in Example 18,substituting(R)—N-(3-chloro-2,2-difluoropropyl)-5-(2-(5-fluoro-2-oxo-1,2-dihydropyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamidefor(R)—N-(3-chloropropyl)-5-(2-(5-fluoro-2-oxo-1,2-dihydropyridin-3-yl)pyrrolidin-1-yl)-N-methylpyrazolo[1,5-a]pyrimidine-3-carboxamidein Step C, and heating at 110° C. for 5 hours, to provide the titleproduct as a pale pink solid (6 mg, 11% yield). MS (apci) m/z=419.3(M+H).

Example 20

(6R)-9-fluoro-13-oxa-2,17,21,22,25-pentaazapentacyclo[17.5.2.0^(2,6).0^(7,12).0^(22,26)]hexacosa-1(25),7,9,11,19(26),20,23-heptaen-18-oneStep A: Preparation of (R)-tert-butyl2-(5-fluoro-2-hydroxyphenyl)pyrrolidine-1-carboxylate

This compound was prepared according to the method described inPreparation A, substituting 3-bromo-5-fluoro-2-methoxypyridine with2-bromo-4-fluorophenyl acetate in Step A (3.2 g, 40% yield). MS (apci)m/z=182.1 (M+H-Boc).

Step B: Preparation of (R)-4-fluoro-2-(pyrrolidin-2-yl)phenolhydrochloride

To a solution of (R)-tert-butyl2-(5-fluoro-2-hydroxyphenyl)pyrrolidine-1-carboxylate (3.2 g, 11.4 mmol)in DCM (20 mL) was added HCl (4 N dioxane, 5.69 mL, 22.7 mmol), and themixture was stirred at ambient temperature for 15 hours. The reactionwas concentrated, and the resulting precipitate was taken up in DCM (15mL) and filtered to afford (R)-4-fluoro-2-(pyrrolidin-2-yl)phenolhydrochloride (1.85 g, 90% yield) as a beige solid. MS (apci) m/z=182.1(M+H).

Step C: Preparation of (R)-ethyl5-(2-(5-fluoro-2-hydroxyphenyl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate

Prepared according to the method described in Preparation B,substituting (R)-4-fluoro-2-(pyrrolidin-2-yl)phenol hydrochloride for(R)-5-fluoro-2-methoxy-3-(pyrrolidin-2-yl)pyridine in Step C. The crudematerial was purified by reverse-phase column chromatography (0-65%acetonitrile/H₂O) to yield the pure product (686 mg, 80% yield). MS(apci) m/z=371.0 (M+H).

Step D: Preparation of (R)-ethyl5-(2-(2-(3-(1,3-dioxoisoindolin-2-yl)propoxy)-5-fluorophenyl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate

A suspension of (R)-ethyl5-(2-(5-fluoro-2-hydroxyphenyl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate(280 mg, 0.756 mmol), 2-(3-bromopropyl)isoindoline-1,3-dione (263 mg,0.983 mmol) and K₂CO₃ (104 mg, 0.756 mmol) in DMF (0.4 mL) was stirredat ambient temperature for 15 hours. The reaction was directly purifiedby reverse-phase column chromatography (5-80% acetonitrile/H₂O) toafford (R)-ethyl5-(2-(2-(3-(1,3-dioxoisoindolin-2-yl)propoxy)-5-fluorophenyl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate(202 mg, 48% yield) as clear oil. MS (apci) m/z=558.0 (M+H).

Step E: Preparation of (R)-ethyl5-(2-(2-(3-aminopropoxy)-5-fluorophenyl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate

(R)-ethyl5-(2-(2-(3-(1,3-dioxoisoindolin-2-yl)propoxy)-5-fluorophenyl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate(200 mg, 0.359 mmol) and hydrazine monohydrate (115 mg, 3.59 mmol) werecombined in MeOH (1 mL) and THF (1 mL) in a sealed vessel and heated at60° C. for 20 minutes. After cooling to ambient temperature, thereaction was concentrated, followed by addition of NaOH (1 N, 2 mL). Themixture was extracted with DCM, and the combined organic extracts weredried (Na₂SO₄), filtered and concentrated to afford (R)-ethyl5-(2-(2-(3-aminopropoxy)-5-fluorophenyl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate(110 mg, 72% yield). MS (apci) m/z=428.2 (M+H).

Step F: Preparation of(6R)-9-fluoro-13-oxa-2,17,21,22,25-pentaazapentacyclo[17.5.2.0^(2,6).0^(7,12).0^(22,26)]hexacosa-1(25),7,9,11,19(26),20,23-heptaen-18-one

(R)-ethyl5-(2-(2-(3-aminopropoxy)-5-fluorophenyl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate(10 mg, 0.023 mmol) and DIEA (8.1 μL, 0.047 mmol) were combined in dryEtOH (0.1 mL) in a sealed vessel and heated at 200° C. overnight. Thereaction was concentrated and purified by reverse-phase columnchromatography (0-70% acetonitrile/H₂O) to afford the title compound(4.5 mg, 50% yield). MS (apci) m/z=382.2 (M+H).

Example 21

(6R)-9-fluoro-13-oxa-2,16,20,21,24-pentaazapentacyclo[16.5.2.0^(2,6).0^(7,12).0^(21,25)]pentacosa-1(24),7,9,11,18(25),19,22-heptaeneStep A: Preparation of(R)-4-fluoro-2-(1-(pyrazolo[1,5-a]pyrimidin-5-yl)pyrrolidin-2-yl)phenol

A mixture of (R)-4-fluoro-2-(pyrrolidin-2-yl)phenol hydrochloride(Example 20, Step B, 1.50 g, 6.89 mmol), DIEA (2.67 g, 20.7 mmol),5-chloropyrazolo[1,5-a]pyrimidine (1.11 g, 7.24 mmol) and isopropanol (1mL) was heated at 120° C. overnight. The reaction was poured into ether(50 mL) and extracted with NaOH (1N aqueous, 3×25 mL). The combinedaqueous extracts were brought to pH 4 with concentrated HCl andextracted with DCM. The combined DCM extracts were filtered throughphase separator paper and concentrated to provide(R)-4-fluoro-2-(1-(pyrazolo[1,5-a]pyrimidin-5-yl)pyrrolidin-2-yl)phenol(1.82 g, 89% yield) as beige solid. MS (apci) m/z=299.4 (M+H).

Step B: Preparation of(R)-5-(2-(5-fluoro-2-hydroxyphenyl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carbaldehyde

After POCl₃ (221 μL, 2.41 mmol) was added drop-wise to a DMF (4 mL)solution of(R)-4-fluoro-2-(1-(pyrazolo[1,5-a]pyrimidin-5-yl)pyrrolidin-2-yl)phenol(600 mg, 2.01 mmol) at ambient temperature, the reaction was stirred for5 minutes before NaOH (804 mg, 10.1 mmol) was introduced. The reactionwas stirred for another 10 minutes before HCl (4 N dioxane, 3 mL) wasadded, followed by DCM (50 mL). After filtering through Celite®, thereaction was concentrated and purified by reverse-phase columnchromatography, eluting with 0-70% acetonitrile/H₂O to provide(R)-5-(2-(5-fluoro-2-hydroxyphenyl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carbaldehyde(524 mg, 80% yield) as beige solid. MS (apci) m/z=327.2 (M+H).

Step C: Preparation of (R)-tert-butyl2-(4-fluoro-2-(1-(3-formylpyrazolo[1,5-a]pyrimidin-5-yl)pyrrolidin-2-yl)phenoxy)ethylcarbamate

A mixture of(R)-5-(2-(5-fluoro-2-hydroxyphenyl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carbaldehyde(159 mg, 0.487 mmol), tert-butyl 2-bromoethylcarbamate (131 mg, 0.585mmol), potassium carbonate (202 mg, 1.46 mmol) and DMF (1 mL) wascombined in a sealed vessel and stirred at ambient temperature overnightand then at 60° C. for 3 hours. After diluting with DCM (20 mL), thereaction was filtered through Celite®, concentrated and purified byreverse-phase column chromatography, eluting with 0-70% acetonitrile/H₂Oto provide (R)-tert-butyl2-(4-fluoro-2-(1-(3-formylpyrazolo[1,5-a]pyrimidin-5-yl)pyrrolidin-2-yl)phenoxy)ethylcarbamate(198 mg, 86.6% yield) as yellowish solid. MS (apci) m/z=370.4 (M+H-Boc).

Step D: Preparation of(R)-5-(2-(2-(2-aminoethoxy)-5-fluorophenyl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carbaldehyde

An HCl (4N dioxane, 80 μl, 0.32 mmol) was added to a DCM (2 mL) solutionof (R)-tert-butyl2-(4-fluoro-2-(1-(3-formylpyrazolo[1,5-a]pyrimidin-5-yl)pyrrolidin-2-yl)phenoxy)ethylcarbamate(198 mg, 0.422 mmol), and the reaction was purged with N₂ and stirred atambient temperature overnight. After removal of solvent, NaOH (5 mL×1N)was introduced and the reaction mixture was extracted with severalportions of DCM in a phase separator tube. The combined organic extractswere concentrated to provide(R)-5-(2-(2-(2-aminoethoxy)-5-fluorophenyl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carbaldehyde (155 mg, 99.5% yield), which was used immediately in the nextstep. MS (apci) m/z=352.3 (M+H—H₂O).

Step E: Preparation of(6R)-9-fluoro-13-oxa-2,16,20,21,24-pentaazapentacyclo[16.5.2.0^(2,6).0^(7,12).0^(21,25)]pentacosa-1(24),7,9,11,18(25),19,22-heptaene

Tetramethylammonium triacetoxyborohydride (46.7 mg, 0.629 mmol) wasadded to a DCM (50 mL) solution of(R)-5-(2-(2-(2-aminoethoxy)-5-fluorophenyl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carbaldehyde(155 mg, 0.420 mmol), and the reaction was stirred at ambienttemperature overnight. The reaction mixture was then diluted with brineand extracted with several portions of DCM in a phase separator tube,and the combined organic extracts were concentrated and purified byreverse-phase column chromatography, eluting with 0-90%acetonitrile-H₂O, to obtain the title product (32 mg, 21.6% yield). MS(apci) m/z=354.2 (M+H).

Example 22

1-[(6R)-9-fluoro-13-oxa-2,16,20,21,24-pentaazapentacyclo[16.5.2.0^(2,6).0^(2,12).0^(21,25)]pentacosa-1(24),7,9,11,18(25),19,22-heptaen-16-yl]ethan-1-one

Acetyl chloride (1.7 mg, 0.021 mmol) was added to a DCM (0.5 mL)solution of(6R)-9-fluoro-13-oxa-2,16,20,21,24-pentaazapentacyclo-[16.5.2.0^(2,6).0^(7,12).0^(21,25)]pentacosa-1(24),7,9,11,18(25),19,22-heptaene(Example 21, 5.0 mg, 0.014 mmol), followed by DIEA (7.4 μL, 0.042 mmol).After stirring at ambient temperature overnight, the reaction wasconcentrated and purified by reverse-phase column chromatography elutingwith 0-80% acetonitrile/H₂O to provide the title product (3.9 mg, 70%yield). MS (apci) m/z=396.2 (M+H).

Example 23

1-[(6R)-9-fluoro-13-oxa-2,16,20,21,24-pentaazapentacyclo[16.5.2.0^(2,6).0^(7,12).0^(21,25)]pentacosa-1(24),7,9,11,18(25),19,22-heptaen-16-yl]-2-hydroxyethan-1-one

To a DCM (0.5 mL) solution of(6R)-9-fluoro-13-oxa-2,16,20,21,24-pentaazapentacyclo-[16.5.2.0^(2,6).0^(7,12).0^(21,25)]pentacosa-1(24),7,9,11,18(25),19,22-heptaene(Example 21, 6 mg, 0.017 mmol) was added 2-chloro-2-oxoethyl acetate(3.5 mg, 0.025 mmol), followed by DIEA (8.9 μL, 0.051 mmol). Thereaction was stirred at ambient temperature overnight, thenconcentrated, and MeOH (0.2 mL) was added followed by sodium hydroxide(6.8 mg, 0.085 mmol). After stirring at ambient temperature for 5 hours,the reaction was diluted with brine and extracted with several portionsof DCM in a phase separator tube. The combined organic extracts wereconcentrated and purified by reverse-phase column chromatography,eluting with 0-70% acetonitrile/H₂O, to provide the title product (3.6mg, 52% yield). MS (apci) m/z=412.5 (M+H).

Example 24

(6R)-9-fluoro-13-oxa-2,17,21,22,25-pentaazapentacyclo[17.5.2.0^(2,6).0^(7,12).0^(22,26)]hexacosa-1(25),7,9,11,19(26),20,23-heptaeneStep A: Preparation of (R)-tert-butyl3-(4-fluoro-2-(1-(3-formylpyrazolo[1,5-a]pyrimidin-5-yl)pyrrolidin-2-yl)phenoxy)propylcarbamate

Prepared according to the method described in Example 21, substitutingtert-butyl 2-bromoethylcarbamate with tert-butyl 3-bromopropylcarbarnatein Step C to afford the desired product (119 mg, 84.5% yield). MS (apci)m/z=384.2 (M+H-Boc).

Step B: Preparation of (R)-tert-butyl3-(4-fluoro-2-(1-(3-(hydroxymethyl)pyrazolo[1,5-a]pyrimidin-5-yl)pyrrolidin-2-yl)phenoxy)propylcarbamate

A solution of (R)-tert-butyl3-(4-fluoro-2-(1-(3-formylpyrazolo[1,5-a]pyrimidin-5-yl)pyrrolidin-2-yl)phenoxy)propylcarbamate(85.0 mg, 0.176 mmol) in MeOH (2 mL) was first cooled to 0° C., thenNaBH₄ (4.04 mg, 0.176 mmol) was introduced, and the reaction was stirredat 0° C. for 1 hour. The reaction was diluted with brine and extractedwith DCM in a phase separator cartridge. The combined organic extractswere concentrated to provide (R)-tert-butyl3-(4-fluoro-2-(1-(3-(hydroxymethyl)pyrazolo[1,5-a]pyrimidin-5-yl)pyrrolidin-2-yl)phenoxy)propylcarbamate(86 mg, 101% yield) as beige solid. MS (apci) m/z=468.1 (M+H—H₂O).

Step C: Preparation of(R)-(5-(2-(2-(3-aminopropoxy)-5-fluorophenyl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidin-3-yl)methanolhydrochloride

(R)-tert-butyl3-(4-fluoro-2-(1-(3-(hydroxymethyl)pyrazolo[1,5-a]pyrimidin-5-yl)pyrrolidine-2-yl)phenoxy)propylcarbamate(80 mg, 0.16 mmol) was dissolved in 2 mL of DCM and treated with HCl (4N in dioxane, 6.0 mg, 0.16 mmol). The reaction was purged with N₂,capped, and stirred at ambient temperature for 18 hours, thenconcentrated to provide(R)-(5-(2-(2-(3-aminopropoxy)-5-fluorophenyl)pyrrolidine-1-yl)pyrazole[1,5-a]pyrimidin-3-yl)methanolhydrochloride (70 mg, 101% yield) as a beige solid. MS (apci) m/z=368.5(M+H—H₂O).

Step D: Preparation of(6R)-9-fluoro-13-oxa-2,17,21,22,25-pentaazapentacyclo-[17.5.2.0^(2,6).0^(7,12).0^(22,26)]hexacosa-1(25),7,9,11,19(26),20,23-heptaene

A mixture of(R)-(5-(2-(2-(3-aminopropoxy)-5-fluorophenyl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidin-3-yl)methanol(50 mg, 0.130 mmol), PS—PPh₃ (0.259 mmol) and perchloromethane (200 mg,1.30 mmol) in DCM (5 mL) was shaken at ambient temperature overnight.The reaction was filtered, concentrated and purified by reverse-phasecolumn chromatography eluting with 0-60% acetonitrile/H₂O to yield thetitle product (27.4 mg, 57.5% yield). MS (apci) m/z=368.1 (M+H).

Example 25

(6R)-9-fluoro-16-methanesulfonyl-13-oxa-2,16,20,21,24-pentaazapentacyclo[16.5.2.0^(2,6).0^(7,12).0^(21,25)]pentacosa-1(24),7,9,11,18(25),19,22-heptaene

To a DCM (0.5 mL) solution of(6R)-9-fluoro-13-oxa-2,16,20,21,24-pentaazapentacyclo-[16.5.2.0^(2,6).0^(7,12).0^(21,25)]pentacosa-1(24),7,9,11,18(25),19,22-heptaene(Example 21, 5 mg, 0.0141 mmol) was added DIEA (2.46 μL, 0.0141 mmol),followed by methanesulfonyl chloride (1.10 μL, 0.0141 mmol). Thereaction was stirred at ambient temperature for 1 hour before MeOH (0.1mL) was added. The reaction was concentrated and purified byreverse-phase column chromatography eluting with 0-80% acetonitrile/H₂Oto provide the title product (3.1 mg, 50.8% yield). MS (apci) m/z=432.3(M+H).

Example 26

2-[(6R)-9-fluoro-13-oxa-2,16,20,21,24-pentaazapentacyclo[16.5.2.0^(2,6).0^(7,12).0^(21,25)]pentacosa-1(24),7,9,11,18(25),19,22-heptaen-16-yl]acetic acid

An IPA (0.1 mL) solution of(6R)-9-fluoro-13-oxa-2,16,20,21,24-pentaazapentacyclo-[16.5.2.0^(2,6).0^(7,12).0^(21,25)]pentacosa-1(24),7,9,11,18(25),19,22-heptaene(Example 21, 5 mg, 0.014 mmol), 2-bromoacetic acid (2.9 mg, 0.021 mmol)and NaOH (1 N, 42 μL, 0.042 mmol) was heated at 60° C. in a sealedvessel overnight, then at 120° C. for 24 hours. After cooling, thereaction mixture was directly purified by reverse-phase columnchromatography eluting with 0-50% acetonitrile/H₂O to afford the titleproduct (3.1 mg, 53% yield). MS (apci) m/z=412.2 (M+H).

Example 27

(6R)-9-fluoro-17-methanesulfonyl-13-oxa-2,17,21,22,25-pentaazapentacyclo[17.5.2.0^(2,6).0^(7,12).0^(22,26)]hexacosa-1(25),7,9,11,19(26),20,23-heptaene

Methanesulfonyl chloride (1.69 μL, 0.0218 mmol) was added to a DCM (0.5mL) solution of(6R)-9-fluoro-13-oxa-2,17,21,22,25-pentaazapentacyclo[17.5.2.0^(2,6).0^(7,12).0^(22,26)]hexacosa-1 (25),7,9,11,19(26),20,23-heptaene (Example 24, 4.0mg, 0.0109 mmol), followed by DIEA (9.48 μL, 0.0544 mmol). The reactionwas stirred at ambient temperature overnight, concentrated and purifiedby reverse-phase column chromatography eluting with 0-80%acetonitrile/H₂O to afford the title compound (2.9 mg, 59.8% yield). MS(apci) m/z=446.3 (M+H).

Example 28

(6R)—N-ethyl-9-fluoro-13-oxa-2,17,21,22,25-pentaazapentacyclo[17.5.2.0^(2,6).0^(7,12).0^(22,26)]hexacosa-1(25),7,9,11,19(26),20,23-heptaene-17-carboxamide

To a DCM (0.5 mL) solution of(6R)-9-fluoro-13-oxa-2,17,21,22,25-pentaazapentacyclo[17.5.2.0^(2,6).0^(7,12).0^(22,26)]hexacosa-1(25),7,9,11,19(26),20,23-heptaene(Example 24, 4 mg, 0.011 mmol) was added isocyanatoethane (1.5 mg, 0.022mmol) followed by DIEA (1.9 μL, 0.011 mmol). The reaction was stirred atambient temperature overnight, then concentrated and purified byreverse-phase column chromatography, eluting with 0-80%acetonitrile/H₂O, to afford the title compound (3.5 mg, 73% yield). MS(apci) m/z=439.1 (M+H).

Example 29

(6R)—N-ethyl-9-fluoro-13-oxa-2,16,20,21,24-pentaazapentacyclo-[16.5.2.0^(2,6).0^(7,12).0^(21,25)]pentacosa-1(24),7,9,11,18(25),19,22-heptaene-16-carboxamide

To a DCM (0.5 mL) solution of(6R)-9-fluoro-13-oxa-2,16,20,21,24-pentaazapentacyclo[16.5.2.0^(2,6).0^(7,12).0^(21,25)]pentacosa-1(24),7,9,11,18(25),19,22-heptaene(Example 21, 5.5 mg, 0.016 mmol) was added isocyanatoethane (1.5 mg,0.022 mmol), followed by DIEA (1.9 μL, 0.011 mmol). After stirring atambient temperature overnight the reaction was concentrated and purifiedby reverse-phase column chromatography, eluting with 0-80%acetonitrile/H₂O to afford the title compound (3.3 mg, 50% yield). MS(apci) m/z=425.4 (M+H).

Example 30

(6S)-9-fluoro-4,13-dioxa-2,11,17,21,22,25-hexaazapentacyclo[17.5.2.0^(2,6).0^(7,12).0^(22,26)]hexacosa-1(25),7(12),8,10,19(26),20,23-heptaene-3,18-dioneStep A: Preparation of(S,E)-N-(2-(tert-butyldimethylsilyloxy)ethylidene)-2-methylpropane-2-sulfinamide

To a solution of (S)-2-methylpropane-2-sulfinimide (3.3 g, 27.2 mmol) inDCM (50 mL) was added 2-(tert-butyldimethylsilyloxy)acetaldehyde (4.98g, 28.6 mmol) followed by anhydrous copper sulfate (8.69 g, 54.5 mmol).The heterogeneous mixture was stirred at ambient temperature for 3 daysand then filtered through Celite®. The filtrate concentrated and theresidue was purified by flash column chromatography, eluting with 10%EtOAc/hexanes, to afford(S,E)-N-(2-(tert-butyldimethylsilyloxy)ethylidene)-2-methylpropane-2-sulfinamide(5.54 g, 73% yield) as a colorless oil. ¹H NMR (CDCl₃) δ 7.96 (m, 1H),4.44 (d, 1H, J=2.7 Hz), 1.11 (s, 9H), 0.82 (s, 9H), 0.00 (s, 6H).

Step B: Preparation of(S)—N—((S)-2-(tert-butyldimethylsilyloxy)-1-(5-fluoro-2-methoxypyridin-3-yl)ethyl)-2-methylpropane-2-sulfinamide

To a solution of n-butyl lithium (10.8 mL, 17.3 mmol, 1.6 M in hexanes)in toluene (100 mL) at −78° C. was added a solution of3-bromo-5-fluoro-2-methoxypyridine (3.27 g, 15.9 mmol) in toluene (5 mL)dropwise, maintaining the internal temperature below −70° C. The mixturewas stirred at −78° C. for 1 hour, then treated with a solution of(S,E)-N-(2-(tert-butyldimethylsilyloxy)ethylidene)-2-methylpropane-2-sulfinamide(4.0 g, 14.4 mmol) in toluene (10 mL) dropwise, maintaining the internaltemperature below −65° C. After stirring at −78° C. for 3 hours themixture was treated with brine (100 mL) and EtOAc (100 mL) and stirredat ambient temperature for 20 minutes. Saturated NaHCO₃ solution (50 mL)was added and the layers were separated. The aqueous layer was extractedwith EtOAc (2×50 mL) and the combined organic phases were washed withbrine (50 mL), dried over Na₂SO₄, filtered and concentrated. The residuewas purified by flash column chromatography, eluting with 10%EtOAc/hexanes to 20% EtOAc/hexanes, to afford (S)—N—((S)-2-(tert-butyldimethylsilyloxy)-1-(5-fluoro-2-methoxypyridin-3-yl)ethyl)-2-methylpropane-2-sulfinamide(1.40 g, 24% yield) mixed with a less polar impurity as a colorless oil.MS (apci) m/z=405.0 (M+H).

Step C: Preparation of(S)-2-amino-2-(5-fluoro-2-methoxypyridin-3-yl)ethanol dihydrochloride

To a solution of(S)—N—((S)-2-(tert-butyldimethylsilyloxy)-1-(5-fluoro-2-methoxypyridin-3-yl)ethyl)-2-methylpropane-2-sulfinamide(1.40 g, 3.46 mmol) in methanol (20 mL) was added 4N HCl/dioxane (8.65mL, 34.6 mmol). The solution was stirred at ambient temperature for 16hours, then concentrated and dried under vacuum to afford(S)-2-amino-2-(5-fluoro-2-methoxypyridin-3-yl)ethanol dihydrochloride asa yellow oil which was used without purification, assuming 100% yield.MS (apci) m/z=186.9 (M+H).

Step D: Preparation of(S)-4-(5-fluoro-2-methoxypyridin-3-yl)oxazolidin-2-one

To a solution of (S)-2-amino-2-(5-fluoro-2-methoxypyridin-3-yl)ethanoldihydrochloride (897 mg, 3.46 mmol) in KOH (10 mL, 24.2 mmol, 2.42 M inwater) was added THF (10 mL). The mixture was cooled to 0° C. andtreated with triphosgene (1.03 g, 3.46 mmol). The mixture was allowed towarm to ambient temperature with stirring over 16 hours then partitionedbetween EtOAc (50 mL) and water (50 mL), and the layers were separated.The aqueous layer was extracted with EtOAc (2×30 mL) and the combinedorganic phases were washed with brine (20 mL), dried over Na₂SO₄,filtered and concentrated. The residue was triturated with Et₂O,filtered and dried under reduced pressure to afford(5)-445-fluoro-2-methoxypyridin-3-yl)oxazolidin-2-one (254 mg, 35%yield) as a white powder. ¹H NMR (CDCl₃) δ 7.98 (m, 1H), 7.44 (m, 1H),5.61 (Br S, 1H), 5.13 (m, 1H), 4.83 (m, 1H), 4.16 (m, 1H), 3.96 (s, 3H).

Step E: Preparation of (S)-ethyl5-(4-(5-fluoro-2-methoxypyridin-3-yl)-2-oxooxazolidin-3-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate

To a solution of (S)-4-(5-fluoro-2-methoxypyridin-3-yl)oxazolidin-2-one(254 mg, 1.20 mmol) in DMF (10 mL) was added sodium hydride (58 mg, 1.44mmol, 60% in mineral oil). The mixture was stirred at ambienttemperature for 20 minutes then treated with ethyl5-chloropyrazolo[1,5-a]pyrimidine-3-carboxylate (270 mg, 1.20 mmol) inone portion. The mixture was stirred for 48 hours then treated withsaturated NH₄Cl solution (30 mL) and extracted with EtOAc (3×10 mL). Thecombined organic phases were washed with water (5×10 mL) and brine (10mL) then dried over Na₂SO₄, filtered and concentrated. The residue waspurified by flash column chromatography, eluting with 20% EtOAc/hexanesto 66% EtOAc/hexanes, to afford (S)-ethyl5-(4-(5-fluoro-2-methoxypyridin-3-yl)-2-oxooxazolidin-3-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate(311 mg, 65% yield) as a white foam. MS (apci) m/z=401.9 (M+H).

Step F: Preparation of(S)-5-(1-(5-fluoro-2-methoxypyridin-3-yl)-2-hydroxyethylamino)pyrazolo[1,5-a]pyrimidine-3-carboxylicacid

To a solution of (S)-ethyl5-(4-(5-fluoro-2-methoxypyridin-3-yl)-2-oxooxazolidin-3-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate(311 mg, 0.77 mmol) in a 1:1:1 mixture of MeOH:THF:H₂O (15 mL) was addedlithium hydroxide monohydrate (97.6 mg, 2.32 mmol). The mixture wasstirred at ambient temperature for 16 hours and then at 50° C. for 19hours, then concentrated to ⅓ volume, diluted with water (30 mL) andacidified to pH 4-5 with 1N HCl. The resulting precipitate was collectedby filtration, washed with water and Et₂O then dried under reducedpressure to afford(S)-5-(1-(5-fluoro-2-methoxypyridin-3-yl)-2-hydroxyethylamino)pyrazolo[1,5-a]pyrimidine-3-carboxylicacid (121 mg, 45% yield) as a white powder. MS (apci) m/z=347.9 (M+H).

Step G: Preparation of(S)—N-(3-chloropropyl)-5-(1-(5-fluoro-2-methoxypyridin-3-yl)-2-hydroxyethylamino)pyrazolo[1,5-a]pyrimidine-3-carboxamide

To a suspension of(S)-5-(1-(5-fluoro-2-methoxypyridin-3-yl-2-hydroxyethylamino)pyrazolo[1,5-a]pyrimidine-3-carboxylicacid (50 mg, 0.14 mmol) in DCM (2 mL) was added HOBt (44 mg, 0.29 mmol)followed by EDCI (83 mg, 0.43 mmol). The heterogeneous mixture wasstirred at ambient temperature for 10 minutes then treated withtriethylamine (100 μL, 0.72 mmol) followed by 3-chloro-propyl aminehydrochloride (56 mg, 0.43 mmol). The mixture was stirred for 2 hours,then DMF (2 mL) was added and stirring was continued for 48 hours. Themixture was partitioned between saturated NH₄Cl solution (20 mL) andEtOAc (20 mL) and the layers were separated. The aqueous layer wasextracted with EtOAc (2×10 mL) and the combined organic phases werewashed with water (5×10 mL) and brine (10 mL), then dried over Na₂SO₄,filtered and concentrated to afford(S)—N-(3-chloropropyl)-5-(1-(5-fluoro-2-methoxypyridin-3-yl)-2-hydroxyethylamino)pyrazolo[1,5-a]pyrimidine-3-carboxamide(60 mg, 99% yield) as a pale yellow foam which was used without furtherpurification. MS (apci) m/z=423.0 (M+H).

Step H: Preparation of(S)—N-(3-chloropropyl)-5-(4-(5-fluoro-2-methoxypyridin-3-yl)-2-oxooxazolidin-3-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

To a solution of(S)—N-(3-chloropropyl)-5-(1-(5-fluoro-2-methoxypyridin-3-yl)-2-hydroxyethylamino)pyrazolo[1,5-a]pyrimidine-3-carboxamide(60 mg, 0.14 mmol) in ACN (2 mL) was added CDI (35 mg, 0.21 mmol). Thesolution was stirred at ambient temperature for 16 hours thenpartitioned between saturated NH₄Cl solution (20 mL) and EtOAc (10 mL)and the layers separated. The aqueous layer was extracted with EtOAc(2×10 mL) and the combined organic phases were washed with brine (10mL), dried over Na₂SO₄, filtered and concentrated. The residue waspurified by flash column chromatography eluting with 1% MeOH/DCM toafford(S)—N-(3-chloropropyl)-5-(4-(5-fluoro-2-methoxypyridin-3-yl)-2-oxooxazolidin-3-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide(37 mg, 58% yield) as a white solid. MS (apci) m/z=449.0 (M+H).

Step I: Preparation of(S)—N-(3-chloropropyl)-5-(4-(5-fluoro-2-oxo-1,2-dihydropyridin-3-yl)-2-oxooxazolidin-3-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

A suspension of(S)—N-(3-chloropropyl)-5-(4-(5-fluoro-2-methoxypyridin-3-yl)-2-oxooxazolidin-3-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide(37 mg, 0.08 mmol) in 4N HCl/dioxane (4 mL) was stirred at 85° C. for 17hours and then at ambient temperature for 48 hours. The resultingsolution was concentrated to ½ volume, transferred to a sealed tube,treated with 4N HCl/dioxane (2 mL) and stirred at 100° C. for 2 hours.The heterogeneous mixture was concentrated, dried under reduced pressureand used directly in the next step, assuming 100% yield. MS (apci)m/z=435.1 (M+H).

Step J: Preparation of(6S)-9-fluoro-4,13-dioxa-2,11,17,21,22,25-hexaazapentacyclo[17.5.2.0^(2,6).0^(7,12).0^(22,26)]hexacosa-1(25),7(12),8,10,19(26),20,23-heptaene-3,18-dione

To a solution of(S)—N-(3-chloropropyl)-5-(4-(5-fluoro-2-oxo-1,2-dihydropyridin-3-yl)-2-oxooxazolidin-3-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (35 mg, 0.08mmol) in DMF (3 mL) was added cesium carbonate (79 mg, 0.24 mmol). Themixture was stirred at 65° C. for 30 minutes then at ambient temperaturefor 48 hours. The mixture was treated with water (30 mL) and extractedwith EtOAc (3×10 mL). The combined organic phases were washed with water(5×10 mL) and brine (10 mL), then dried over Na₂SO₄, filtered andconcentrated. The residue was purified via flash column chromatographyeluting with 2% MeOH/DCM to afford the title compound (13 mg, 41% yield)as an amorphous white solid. MS (apci) m/z=399.2 (M+H).

Example 31

(6S)-9-fluoro-4,13-dioxa-2,11,16,20,21,24-hexaazapentacyclo[16.5.2.0^(2,6).0^(7,12).0^(21,25)]pentacosa-1(24),7(12),8,10,18(25),19,22-heptaene-3,17-dioneStep A: Preparation of 5-hydroxypyrazolo[1,5-a]pyrimidine-3-carboxylicacid

To a solution of ethyl 5-hydroxypyrazolo[1,5-a]pyrimidine-3-carboxylate(Preparation B, Step A; 2.0 g, 9.65 mmol) in a 2:1 mixture of THF:MeOH,(40 mL) was added lithium hydroxide monohydrate (29 mL, 29.0 mmol, 1.0 Min water). The solution was stirred at reflux for 16 hours then cooledand concentrated. The residue was dissolved in water (100 mL) andacidified with 6M HCl. The resulting white precipitate was collected byfiltration and washed with water and Et₂O, then dried under reducedpressure to afford 5-hydroxypyrazolo[1,5-a]pyrimidine-3-carboxylic acid(1.18 g, 68% yield) as a white solid. ¹H NMR (d₆-DMSO) δ 8.50 (d, 2H,J=7.7 Hz), 8.02 (s, 2H), 6.07 (d, 2H, J=8.2 Hz).

Step B: Preparation of 5-chloropyrazolo[1,5-a]pyrimidine-3-carbonylchloride

To a suspension of 5-hydroxypyrazolo[1,5-a]pyrimidine-3-carboxylic acid(1.18 g, 6.59 mmol) in DMF (10 mL) at 0° C. was added thionyl chloride(10 mL) dropwise over 5 minutes. The mixture was warmed to ambienttemperature, then stirred at 60° C. for 16 hours. The cooled solutionwas purged with N₂ for 20 minutes then diluted with 50% EtOAc/hexanes(100 mL) and stirred vigorously for 30 minutes. The organic phase wasdecanted, treated with Na₂CO₃ and activated carbon, stirred for 5minutes then filtered through Centel) and concentrated. The residue wasdissolved in toluene (100 mL), treated with activated carbon andfiltered through Celite® again. The filtrate was concentrated and driedunder reduced pressure to afford5-chloropyrazolo[1,5-a]pyrimidine-3-carbonyl chloride (800 mg, 56%yield) as a cream-colored solid. ¹H NMR (CDCl₃) δ 8.70 (m, 1H), 8.66 (s,1H), 7.16 (m, 1H).

Step C: Preparation of5-chloro-N-(2-chloroethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

To a suspension of 5-chloropyrazolo[1,5-a]pyrimidine-3-carbonyl chloride(284 mg, 1.31 mmol) in DCM (10 mL) was added DIEA (1.14 mL, 6.57 mmol).The solution was cooled to 0° C., then treated with 2-chloroethylaminehydrochloride (183 mg, 1.58 mmol) and stirred for 1 hour. The mixturewas partitioned between water (30 mL) and DCM (30 mL) and the layerswere separated. The aqueous layer was extracted with DCM (2×20 mL) andthe combined organic phases were washed with brine (20 mL), dried overNa₂SO₄, filtered and concentrated to afford5-chloro-N-(2-chloroethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (290mg, 85% yield) as a beige solid. MS (apci) m/z=258.9 (M+H).

Step D: Preparation of(S)—N-(2-chloroethyl)-5-(4-(5-fluoro-2-methoxypyridin-3-yl)-2-oxooxazolidin-3-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

To a solution of (S)-4-(5-fluoro-2-methoxypyridin-3-yl)oxazolidin-2-one(prepared according to Example 30; 50 mg, 0.236 mmol) in DMF (1 mL) wasadded sodium hydride (11 mg, 0.28 mmol, 60% in mineral oil). The mixturewas stirred at ambient temperature for 20 minutes, then treated with5-chloro-N-(2-chloroethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (61mg, 0.236 mmol). The mixture was stirred at 16 hours, and then treatedwith saturated NH₄Cl solution (10 mL) and water (20 mL). The resultingprecipitate was collected by filtration, washed with water and Et₂O,then dried under reduced pressure to afford(S)—N-(2-chloroethyl)-5-(4-(5-fluoro-2-methoxypyridin-3-yl)-2-oxooxazolidin-3-yl) pyrazolo[1,5-a]pyrimidine-3-carboxamide (83 mg, 81%yield) as a beige solid. MS (apci) m/z=434.9 (M+H).

Step E: Preparation of(S)—N-(2-chloroethyl)-5-(4-(5-fluoro-2-oxo-1,2-dihydropyridin-3-yl)-2-oxooxazolidin-3-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

A suspension of(S)—N-(2-chloroethyl)-5-(4-(5-fluoro-2-methoxypyridin-3-yl)-2-oxooxazolidin-3-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide(80 mg, 0.18 mmol) in 5-6N HCl/IPA (2.5 mL) was warmed to 90° C. in asealed tube for 1.5 hours. The cooled mixture was filtered and thefiltrate was concentrated. The residue was concentrated twice from Et₂Oand dried under reduced pressure to afford(S)—N-(2-chloroethyl)-5-(4-(5-fluoro-2-oxo-1,2-dihydropyridin-3-yl)-2-oxooxazolidin-3-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide(63 mg, 82% yield) as a beige solid. MS (apci) m/z=421.0 (M+H).

Step F: Preparation of(6S)-9-fluoro-4,13-dioxa-2,11,16,20,21,24-hexaazapentacyclo[16.5.2.0^(2,6).0^(7,12).0^(21,25)]pentacosa-1(24),7(12),8,10,18(25),19,22-heptaene-3,17-dione

Prepared according to the method of Example 30, Step J, using(S)—N-(2-chloroethyl)-5-(4-(5-fluoro-2-oxo-1,2-dihydropyridin-3-yl)-2-oxooxazolidin-3-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamidein place of(S)—N-(3-chloropropyl)-5-(4-(5-fluoro-2-oxo-1,2-dihydropyridin-3-yl)-2-oxooxazolidin-3-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamideto afford the title compound (14 mg, 24% yield) as a white solid. MS(apci) m/z=385.1 (M+H).

Example 32

(6R)-9-fluoro-2,11,16,20,21,24-hexaazapentacyclo[16.5.2.0^(2,6).0^(7,12).0^(21,25)]pentacosa-1(24),7,9,11,18(25),19,22-heptaen-17-oneStep A: Preparation of (R)-ethyl5-(2-(2-(3-((tert-butoxycarbonyl)amino)prop-1-yn-1-yl)-5-fluoropyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate

(R)-ethyl 5-(2-(2-chloro-5-fluoropyridin-3-yl)pyrrolidin-1-yl)pyrazolopyrimidine-3-carboxylate (Example 12, Step C; 153 mg, 0.392 mmol) in DMF(2 mL) was added tert-butyl prop-2-ynylcarbamate (122 mg, 0.785 mmol),copper(I) iodide (11 mg, 0.0578 mmol), triphenylphosphine (82.4 mg,0.314 mmol), di-triphenylphosphine palladium(II) chloride (116 mg, 0.165mmol), and diisopropylamine (99.3 mg, 0.981 mmol). The reaction mixturewas sealed and heated to 95° C. for 8 hours, then cooled to ambienttemperature and concentrated under reduced pressure. The residue waspurified by silica column chromatography, eluting with 33% EtOAc/Hexanesto afford the final product mixed with Ph₃P (160 mg, 80.2% yield). MS(apci) m/z=508.9 (M+H).

Step B: Preparation of (R)-ethyl5-(2-(2-(3-aminopropyl)-5-fluoropyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate

To (R)-ethyl5-(2-(2-(3-(tert-butoxycarbonylamino)prop-1-ynyl)-5-fluoropyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate(160 mg, 0.315 mmol) in MeOH (10 mL) was added dihydroxypalladium (101mg, 0.144 mmol). The reaction mixture was stirred under a hydrogenballoon for 6 hours, then filtered through a pad of Celitek and washedwith MeOH (30 mL). The filtrate was concentrated and the resultantresidue was treated with 4 M HCl in dioxane (3 mL). After stirring for30 minutes, the solution was concentrated to afford the product as anHCl salt (140 mg, 108% yield). MS (apci) m/z=413.0 (M+H).

Step C: Preparation of(R)-5-(2-(2-(3-aminopropyl)-5-fluoropyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylicacid

To (R)-ethyl5-(2-(2-(3-aminopropyl)-5-fluoropyridin-3-yl)pyrrolidin-1-yl)pyrazolopyrimidine-3-carboxylate hydrochloride (160 mg, 0.356 mmol) in THF/MeOH(2 mL/1 mL) was added lithium hydroxide (1.1 mL, 2.20 mmol). Thereaction mixture was heated to 70° C. for 5 hours, then concentratedunder reduced pressure. Water (10 mL) was added and the mixture washedwith Et₂O (2×5 mL), then neutralized with HCl (1M) to pH=4. The aqueoussolution was extracted with DCM (2×10 mL). The organic extract was driedwith Na₂SO₄, filtered and concentrated under reduced pressure to givethe crude desired product (16.0 mg, 11.7% yield). MS (apci) m/z=385.0(M+H).

Step D: Preparation of(6R)-9-fluoro-2,11,16,20,21,24-hexaazapentacyclo[16.5.2.0^(2,6).0^(7,12).0^(21,25)]pentacosa-1(24),7,9,11,18(25),19,22-heptaen-17-one

To(R)-5-(2-(2-(3-aminopropyl)-5-fluoropyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylicacid (16 mg, 0.042 mmol) in DMF (5 mL) was added HATU (63 mg, 0.17 mmol)and N-ethyl-N-isopropylpropan-2-amine (22 mg, 0.17 mmol). The reactionmixture was stirred for 3 hours and concentrated under reduced pressure.The crude residue was purified by silica column chromatography using100% EtOAc to afford the title compound (6.0 mg, 39% yield). MS (apci)m/z=367.3 (M+H).

Example 33

(6R)-9-fluoro-15-methyl-2,11,16,20,21,24-hexaazapentacyclo[16.5.2.0^(2,6).0^(7,12).0^(21,25)]pentacosa-1(24),7,9,11,18(25),19,22-heptaen-17-one

Prepared according to the method of Example 37, substituting tert-butyl2-methylbut-3-yn-2-ylcarbamate with tert-butyl but-3-yn-2-ylcarbamate inStep B to afford the title compound as a 1:1 mixture of diastereomers.MS (apci) m/z=381.2 (M+H).

Example 34

(6R,13R)-9-fluoro-13-methyl-2,11,15,19,20,23-hexaazapentacyclo[15.5.2.1^(7,11).0^(2,6).0^(20,24)]pentacosa-1(23),7,9,17(24),18,21-hexaene-16,25-dioneStep A: Preparation of (R)-methyl5-(2-(5-fluoro-2-hydroxypyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate

To a suspension of(R)-5-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylicacid (Preparation B; 5.01 g, 14.0 mmol) in MeOH (150 mL) was addeddropwise TMSCHN₂ (8.41 mL, 16.8 mmol). The reaction was stirred for 30minutes, and then quenched with 1 mL of acetic acid. The solvent wasremoved under reduced pressure and the residue was dried under highvacuum to give the crude methyl ester. To the crude methyl ester wasadded 4N HCl in dioxane (100 mL) and the reaction was sealed and heatedto 90° C. for 2 hours. The reaction mixture was concentrated underreduced pressure and the residue was dissolved in DCM (100 mL) andwashed with saturated NaHCO₃ (40 mL). The organic layer was dried overNa₂SO₄, filtered and concentrated under reduced pressure to give thecrude desired product (4.67 g, 93.2% yield). MS (apci) m/z=357.9 (M+H).

Step B: Preparation of methyl5-((R)-2-(1-((S)-3-(1,3-dioxoisoindolin-2-yl)-2-methylpropyl)-5-fluoro-2-oxo-1,2-dihydropyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate

To a solution of (R)-methyl5-(2-(5-fluoro-2-hydroxypyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate(202 mg, 0.565 mmol) in DMF (5 mL) was added lithium hydride (22.5 mg,2.83 mmol) and (R)-2-(3-bromo-2-methylpropyl)isoindoline-1,3-dione(prepared according to the procedure described in Euro. J. Med. Chem.2000, 147-156) (239 mg, 0.848 mmol). The reaction was stirred for 2hours at 70° C., and then cooled to ambient temperature. The reactionmixture was diluted with EtOAc (20 mL) and washed with water (2×10 mL).The organic layer was dried with Na₂SO₄, filtered and concentrated underreduced pressure. The residue was purified by silica columnchromatography, eluting with 66% EtOAc/Hexanes to afford the product(110 mg, 34.8% yield). MS (apci) m/z=559.0 (M+H).

Step C: Preparation of methyl5-((R)-2-(14R)-3-amino-2-methylpropyl)-5-fluoro-2-oxo-1,2-dihydropyridin-3-yl)pyrrolidin-1-yl)pyrazolopyrimidine-3-carboxylate

To a solution of methyl5-((R)-2-(1-((S)-3-(1,3-dioxoisoindolin-2-yl)-2-methylpropyl)-5-fluoro-2-oxo-1,2-dihydropyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate(110 mg, 0.197 mmol) in McOH/THF (3 mL/3 mL) was added hydrazine (31.6mg, 0.985 mmol). The reaction was stirred for 14 hours at 50° C. Aftercooling, the reaction mixture was concentrated and the resulting residuewas diluted with EtOAc (20 mL) and washed with saturated aqueous NaHCO₃(5 mL), water (2×5 mL) and brine (5 mL). The organic layer was driedwith Na₂SO₄, filtered and concentrated under reduced pressure. Theresidue was purified by silica column chromatography, eluting withEtOAc/MeOH/NH₄OH 10:1:0.1 to give the desired product (65 mg, 77%yield). MS (apci) m/z=429.2 (M+H).

Step D: Preparation of(6R,13R)-9-fluoro-13-methyl-2,11,15,19,20,23-hexaazapentacyclo[15.5.2.1^(7,11).0^(2,6).0^(20,24)]pentacosa-1(23),7,9,17(24),18,21-hexaene-16,25-dione

To a solution of methyl5-((R)-2-(1-((R)-3-amino-2-methylpropyl)-5-fluoro-2-oxo-1,2-dihydropyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate(65 mg, 0.15 mmol) in THF/MeOH (6 mL/2 mL) was added lithium hydroxide(455 μL, 0.91 mmol). The reaction was stirred at 70° C. for 3 hours,then quenched with hydrogen chloride (910 μL, 0.91 mmol). The solventwas removed under reduced pressure and the residue was dried under highvacuum. To the resulting crude residue was added DMF (10 mL), HATU (115mg, 0.30 mmol) and N-ethyl-N-isopropylpropan-2-amine (78 mg, 0.61 mmol).The reaction was stirred for 3 hours, and the solvent was removed underreduced pressure. The residue was purified by silica columnchromatography, eluting with 10% MeOH/EtOAc to afford the title compound(6.0 mg, 10% yield). MS (apci) m/z=397.3 (M+H).

Example 35

(6R,13S)-9-fluoro-13-methyl-2,11,15,19,20,23-hexaazapentacyclo[15.5.2.1^(7,11).0^(2,6).0^(20,24)]pentacosa-1(23),7,9,17(24),18,21-hexaene-16,25-dione

Prepared according to the method of Example 34, substituting(S)-2-(3-bromo-2-methylpropyl)isoindoline-1,3-dione (prepared accordingto the procedure described in Euro. J. Med. Chem. 2000, 147-156) for(R)-2-(3-bromo-2-methylpropyl)isoindoline-1,3-dione in Step B. MS(apci)=397.3 (M+H).

Example 36

(6R)-9-fluoro-15,15-dimethyl-13-oxa-2,11,17,21,22,25-hexaazapentacyclo[17.5.2.0^(2,6).0^(7,12).0^(22,26)]hexacosa-1(25),7,9,11,19(26),20,23-heptaen-18-one

Prepared according to the procedure for Example 3, substituting3-amino-2,2-dimethylpropan-1-ol for 3-aminopropan-1-ol in Step A. MS(apci) m/z=411.2 (M+H).

Example 37

(6R)-9-fluoro-15,15-dimethyl-2,11,16,20,21,24-hexaazapentacyclo[16.5.2.0^(2,6).0^(7,12).0^(21,25)]pentacosa-1(24),7,9,11,18(25),19,22-heptaen-17-oneStep A: Preparation of (R)-methyl5-(2-(5-fluoro-2-(trifluoromethyl-sulfonyloxy)pyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate

To a solution of (R)-methyl5-(2-(5-fluoro-2-hydroxypyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate(Prepared according to Example 34, Step A; 2.31 g, 6.46 mmol) in DMF (20mL) was added1,1,1-trifluoro-N-phenyl-N-(trifluoromethyl-sulfonyl)methanesulfonamide(2.54 g, 7.11 mmol) and triethylamine (0.785 g, 7.76 mmol). The reactionwas stirred for 18 hours. The solvent was removed under reduced pressureand the residue was purified by silica column chromatography, elutingwith 33% EtOAc/Hexanes to afford the desired product (2.36 g, 74.6%yield). MS (apci) m/z=490.0 (M+H).

Step B: Preparation of (R)-methyl5-(2-(2-(3-(tert-butoxycarbonylamino)-3-methylbutyl)-5-fluoropyridin-3-yl)pyrrolidin-1-yl)pyrazolopyrimidine-3-carboxylate

To (R)-methyl5-(2-(5-fluoro-2-(trifluoromethylsulfonyloxy)pyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate(503 mg, 1.03 mmol) in DMF (2 mL) was added tert-butyl2-methylbut-3-yn-2-ylcarbamate (377 mg, 2.06 mmol), copper(I) iodide(39.1 mg, 0.206 mmol), di-triphenylphosphine palladium(II) chloride (144mg, 0.206 mmol), diisopropylamine (260 mg, 2.57 mmol). The reactionmixture was sealed and heated to 65° C. for 8 hours. The solvent wasremoved under reduced pressure. The residue was purified by silicacolumn chromatography, eluting with 66% EtOAc/Hexanes to give (R)-methyl5-(2-(2-(3-(tert-butoxycarbonylamino)-3-methylbut-1-ynyl)-5-fluoropyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylatemixed with Ph₃P, which was immediately hydrogenated usingdihydroxypalladium on carbon (200 mg, 0.285 mmol) in MeOH (20 mL) undera H₂ balloon for 15 hours. After filtering through a pad of Celite® andwashing with MeOH, the filtrate was concentrated under reduced pressureand purified by silica column chromatography, eluting with 66%EtOAc/Hexanes to afford the product (166 mg, 30.7% yield). MS(apci)=527.1 (M+H).

Step C: Preparation of(6R)-9-fluoro-15,15-dimethyl-2,11,16,20,21,24-hexaazapentacyclo[16.5.2.0^(2,6).0^(7,12).0^(21,25)]pentacosa-1(24),7,9,11,18(25),19,22-heptaen-17-one

To (R)-methyl5-(2-(2-(3-(tert-butoxycarbonylamino)-3-methylbutyl)-5-fluoropyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate(166 mg, 0.315 mmol) in THF/MeOH (3 mL/1 mL) was added lithium hydroxide(946 μL, 1.89 mmol). The reaction vessel was sealed and heated to 70° C.for 3 hours. The reaction mixture was then dried under reduced pressureand HCl (4 mL, 4M in dioxane) was added. The reaction mixture wasstirred for one hour, then the solvent was removed and the residue wasdried under high vacuum for two hours. To the residue was then added DMF(8 mL), HOBT-H₂O (96.5 mg, 0.630 mmol), EDCI (121 mg, 0.630 mmol) andtriethylamine (159 mg, 1.58 mmol). The reaction mixture was stirred at45° C. for 18 hours, then concentrated under vacuum. The residue waspurified by silica column chromatography eluting with 5% MeOH/DCM toafford the title compound (60.0 mg, 48.3% yield). MS (apci) m/z=395.1(M+H).

Example 38

(6R)-9-fluoro-13-oxa-2,11,16,17,21,25,26,29-octaazahexacyclo[21.5.2.0^(2,6).0^(7,12).0^(16,20).0^(26,30)]triaconta-1(29),7,9,11,17,19,23(30),24,27-nonaen-22-one Step A: Preparation of1-(2-(tert-butyldiphenylsilyloxy)ethyl)-1H-pyrazol-5-amine

To a suspension of 2-(5-amino-1H-pyrazol-1-yl)ethanol (2.07 g, 16.0mmol) and 1H-imidazole (5.43 g, 79.8 mmol) in DMF (10 mL) was addeddropwise tert-butylchlorodiphenylsilane (4.96 mL, 19.1 mmol). Thereaction was stirred for 15 hours. The solvent was removed under reducedpressure and the residue was diluted with DCM (40 mL). The organic layerwas washed with 1N HCl (10 mL), water (10 mL) and brine (10 mL), thenconcentrated to give crude desired product (5.62 g, 96.4% yield), whichwas used in the next step without purification.

Step B: Preparation of(R)—N-(1-(2-(tert-butyldiphenylsilyloxy)ethyl)-1H-pyrazol-5-yl)-5-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5a]pyrimidine-3-carboxamide

To a suspension of(R)-5-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylicacid (220 mg, 0.616 mmol) in DMF (5 mL) was added dropwise2,4,6-trichlorobenzoyl chloride (106 μL, 0.677 mmol) and triethylamine(81.0 mg, 0.800 mmol). The reaction was stirred for 2 hours, and1-(2-(tert-butyldiphenylsilyloxy)ethyl)-1H-pyrazol-5-amine (338 mg,0.923 mmol) was added to the reaction mixture. The reaction was heatedto 60° C. for 3 hours and then cooled to ambient temperature. Thesolvent was removed under reduced pressure and the residue was purifiedby silica column chromatography to afford the desired product (201 mg,46.3% yield). MS (apci) m/z=705.1 (M+H).

Step C: Preparation of(6R)-9-fluoro-13-oxa-2,11,16,17,21,25,26,29-octaazahexacyclo[21.5.2.0^(2,6).0^(7,12).0^(16,20).0^(26,30)]triaconta-1(29),7,9,11,17,19,23(30),24,27-nonaen-22-one

A suspension of(R)—N-(1-(2-(tert-butyldiphenylsilyloxy)ethyl)-1H-pyrazol-5-yl)-5-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)pyrazolopyrimidine-3-carboxamide (201 mg, 0.285 mmol) in 4M HCl in dioxane (6mL) was sealed and heated to 100° C. for fours hours. The reactionmixture was cooled to ambient temperature and concentrated under reducedpressure. The residue was diluted with DCM (20 mL) and washed withsaturated NaHCO₃ (5 mL), water (5 mL) and brine (5 mL). The organiclayer was concentrated to give crude(R)-5-(2-(5-fluoro-2-hydroxypyridin-3-yl)pyrrolidin-1-yl)-N-(1-(2-hydroxyethyl)-1H-pyrazol-5-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide,to which was added THF (20 mL), DEAD (53.9 μL, 0.342 mmol) andtriphenylphosphine (89.8 mg, 0.342 mmol). The reaction mixture wasstirred for 18 hours, then concentrated under vacuum. The residue waspurified by silica column chromatography, eluting with 10% MeOH/DCM toafford the title compound (1.8 mg, 1.5% yield). MS (apci) m/z=435.3(M+H).

Example 39

(6R)-9-fluoro-13-oxa-2,11,19,21,25,26,29-heptaazahexacyclo[21.5.2.0^(2,6).0^(7,12).0^(15,20).0^(26,30)]triaconta-1(29),7,9,11,15(20),16,18,23(30),24,27-decaen-22-oneStep A: Preparation of3-((tert-butyldiphenylsilyloxy)methyl)pyridin-2-amine

To a suspension of (2-aminopyridin-3-yl)methanol (2.19 g, 17.6 mmol) and1H-imidazole (6.00 g, 88.2 mmol) in DMF (10 mL) was added dropwisetert-butylchlorodiphenylsilane (5.49 mL, 21.2 mmol). The reaction wasstirred for 15 hours. The solvent was removed under reduced pressure andthe residue was diluted with DCM (4 0 mL). The organic layer was washedwith 1N HCl (10 mL), water (10 mL) and brine (10 mL) and thenconcentrated to give crude product (6.03 g, 94.3% yield). MS (apci)m/z=363.1 (M+H).

Step B: Preparation of(6R)-9-fluoro-13-oxa-2,11,19,21,25,26,29-heptaazahexacyclo[21.5.2.0^(2,6).0^(7,12).0^(15,20).0^(26,30)]triaconta-1(29),7,9,11,15(20),16,18,23(30),24,27-decaen-22-one

To a suspension of(R)-5-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylicacid (303 mg, 0.848 mmol) in DMF (5 mL) was added triethylamine (103 mg,1.02 mmol), followed by dropwise addition of 2,4,6-trichlorobenzoylchloride (227 mg, 0.933 mmol). The reaction was stirred for two hours.3-((tert-Butyldiphenylsilyloxy)methyl)pyridin-2-amine (369 mg, 1.02mmol) was added and the reaction mixture was heated to 60° C. for 5hours. The solvent was removed under reduced pressure and the residuewas purified by silica column chromatography, eluting with 10% MeOH/DCMto give(R)—N-(3-((tert-butyldiphenylsilyloxy)methyl)pyridin-2-yl)-5-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide,to which was added THF (5 mL) and TBAF (848 μL, 0.848 mmol). Thereaction mixture was stirred for one hour, then quenched with saturatedNH₄Cl (1 mL) and then concentrated under reduced pressure to give crude(R)-5-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)-N-(3-(hydroxymethyl)pyridin-2-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide,to which was added HCl (4M in dioxane, 5 mL). The reaction mixture wassealed and heated to 100° C. for four hours. The reaction mixture wascooled to ambient temperature and concentrated under reduced pressure.The residue was diluted with DCM (20 mL) and the organic layer waswashed with saturated NaHCO₃ (5 mL), water (5 mL) and brine (5 mL). Theorganic layer was concentrated under reduced pressure to give crude(R)—N-(3-(chloromethyl)pyridin-2-yl)-5-(2-(5-fluoro-2-hydroxypyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide,to which was added DMF (10 mL) and Cs₂CO₃ (276 mg, 0.848 mmol). Thereaction mixture was heated to 60° C. for 4 hours, then cooled toambient temperature and concentrated under reduced pressure. The residuewas purified by silica column chromatography, eluting with 10% MeOH/DCMto afford the title compound (8.0 mg, 2.2% yield). MS (apci) m/z=432.3(M+H).

Example 40

(6R)-9-fluoro-13,13-dimethyl-2,11,15,19,20,23-hexaazapentacyclo[15.5.2.1^(7,11).0^(2,6).0^(20,24)]pentacosa-1(23),7,9,17(24),18,21-hexaene-16,25-dioneStep A: Preparation of 3-bromo-2,2-dimethylpropan-1-amine hydrobromide

A mixture of 2-(3-bromo-2,2-dimethylpropyl)isoindoline-1,3-dione (1.00g, 3.38 mmol) in 48% aqueous HBr (10 mL) was refluxed for 18 hours. Thereaction mixture was cooled to ambient temperature and the solids formedwere filtered off. The filtrate was concentrated under reduced pressureto give the crude material that was azeotroped with toluene (3×)followed by acetonitrile until solids formed. The crude material wastriturated with ether and dried under reduced pressure to afford3-bromo-2,2-dimethylpropan-1-amine hydrobromide (0.816 g, 3.07 mmol,91.0% yield) (confirmed by ¹H-NMR and posAPCI-MS). The isolated productwas used directly without further purification.

Step B: Preparation of(R)—N-(3-bromo-2,2-dimethylpropyl)-5-(2-(5-fluoro-2-hydroxypyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

To a solution of(R)-5-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylicacid (Preparation B; 150 mg, 0.420 mmol), EDCI (88.5 mg, 0.462 mmol),and HOBT-H₂O (70.7 mg, 0.462 mmol) in DMF (10 mL) was added3-bromo-2,2-dimethylpropan-1-amine hydrobromide (124 mg, 0.504 mmol)followed by triethylamine (55.2 mg, 0.546 mmol). The reaction wasstirred for 18 hours. The solvent was removed under reduced pressure andthe residue was purified by silica column chromatography, eluting with50% EtOAc/Hexanes to provide(R)—N-(3-bromo-2,2-dimethylpropyl)-5-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide(180 mg), to which was added HCl (5 mL, 4M in dioxane). The reaction wassealed and heated to 90° C. for 2 hours. The solvent was removed underreduced pressure and the residue was purified by silica columnchromatography, eluting with 20% Hexanes/EtOAc to provide the desiredproduct (130 mg, 63% yield).

Step C: Preparation of(6R)-9-fluoro-13,13-dimethyl-2,11,15,19,20,23-hexaazapentacyclo[15.5.2.1^(7,11).0^(2,6).0^(20,24)]pentacosa-1(23),7,9,17(24),18,21-hexaene-16,25-dione

To a solution of(R)—N-(3-bromo-2,2-dimethylpropyl)-5-(2-(5-fluoro-2-hydroxypyridin-3-yl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide(30 mg, 0.061 mmol) in THF (5 mL) was added drop-wise potassium2-methylpropan-2-olate (153 μL, 0.15 mmol). The reaction was heated at50° C. for two hours. The solvent was removed under reduced pressure andthe residue was purified by silica column chromatography, eluting with10% MeOH/DCM to provide the title compound (15 mg, 60% yield). MS (apci)m/z=411.0 (M+H).

Example 41

(4R,6R,15S)-9-fluoro-4,15-dihydroxy-13-oxa-2,17,21,22,25-pentaazapentacyclo[17.5.2.0^(2,6).0^(7,12).0^(22,26)]hexacosa-1(25),7(12),8,10,19(26),20,23-heptaen-18-oneStep A: Preparation ofN—((S)-3-chloro-2-hydroxypropyl)-5-((R)-2-(5-fluoro-2-hydroxyphenyl)-4-hydroxypyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

To a suspension of(R)-5-(2-(5-fluoro-2-hydroxyphenyl)-4-hydroxypyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylicacid (Preparation D; 0.0339 g, 0.0946 mmol) and HATU (0.0540 g, 0.142mmol) in DMF (0.5 mL) at 0° C. was added (S)-1-amino-3-chloropropan-2-olhydrochloride (Example 19, Step A; 0.0155 g, 0.142 mmol; preparedaccording to the method described in Org. Process Res. Dev. 2003, vol.7, p. 533) and N,N-diisopropylethylamine (0.0494 mL, 0.284 mmol). Theresulting mixture was warmed to ambient temperature and stirred for 18hours. The reaction mixture was diluted with EtOAc (10 mL), washed withbrine, dried over MgSO₄, filtered, and concentrated under reducedpressure to give the crude material which was purified by silica columnchromatography, eluting with 0-20% MeOH/DCM to affordN-((5)-3-chloro-2-hydroxypropyl)-5-((R)-2-(5-fluoro-2-hydroxyphenyl)-4-hydroxypyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide(as a mixture of cis and trans isomers, 0.0407 g, 82.2% yield, 86%purity). LC/MS (ES+APCI) m/z=448.1 (M−H).

Step B: Preparation of (4R,6R,15S)-9-fluoro-4,15-dihydroxy-13-oxa-2,17,21,22,25-pentaazapentacyclo[17.5.2.0^(2,6).0^(7,12).0^(22,26)]hexacosa-1(25),7(12),8,10,19(26),20,23-heptaen-18-one

A mixture ofN—((S)-3-chloro-2-hydroxypropyl)-5-((R)-2-(5-fluoro-2-hydroxyphenyl)-4-hydroxypyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide(0.0407 g, 0.0778 mmol) and Cs₂CO₃ (0.127 g, 0.389 mmol) in DMF (3.6 mL)was heated at 85° C. for 30 minutes. The reaction mixture was cooled toambient temperature and filtered. The filtrate was concentrated underreduced pressure to give the crude material which was purified by silicacolumn chromatography, eluting with 0-20% MeOH/EtOAc to afford the crudeproduct. The crude material was purified using chiral columnchromatography (Chiral Tech OD-H column, 20% EtOH in hexanes). Isolationof the material having a retention time of about 21.8 minutes affordedthe title compound (0.0052 g, 16.2% yield). The stereochemistry of thetitle compound was confirmed by ¹H-NMR nOe experiment. LC/MS (ES+APCI)m/z=414.1 (M+H).

Example 41-B

(4R,6S,15S)-9-fluoro-4,15-dihydroxy-13-oxa-2,17,21,22,25-pentaazapentacyclo[17.5.2.0^(2,6).0^(7,12).0^(22,26)]hexacosa-1(25),7(12),8,10,19(26),20,23-heptaen-18-one

The title compound was isolated during chiral separation reported inExample 41 from fractions having a retention time of about 30.6 minutes,to provide 5.4 mg (16.8% yield) of the compound which may have beenisolated along with the enantiomer and/or one or more diastercomers. Thestereochemistry of the title compound was confirmed by ¹H-NMR nOeexperiment), LC/MS (ES+APCI) m/z=414.1 (M+H).

Example 42

(4R,6R)-9-fluoro-4-hydroxy-13-oxa-2,17,21,22,25-pentaazapentacyclo[17.5.2.0^(2,6).0^(7,12).0^(22,26)]hexacosa-1(25),7(12),8,10,19(26),20,23-heptaen-18-one

The title compound was prepared according to the method of Example 41,substituting 3-chloropropan-1-amine hydrochloride for(5)-1-amino-3-chloropropan-2-ol hydrochloride in Step A: 13.8 mg (16%yield; Chiral Tech OD-H column, 20% EtOH in hexanes, retention timeabout 17.2 minutes). The stereochemistry of the title compound wasconfirmed by ¹H-NMR nOe experiment. LC/MS (ES-hAPCI) m/z=398.1 (M+H).

Example 42-B

(4R,6S)-9-fluoro-4-hydroxy-13-oxa-2,17,21,22,25-pentaazapentacyclo[17.5.2.0^(2,6).0^(7,12).0^(22,26)]hexacosa-1(25),7(12),8,10,19(26),20,23-heptaen-18-one

The title compound was prepared during the chiral separation reported inExample 42 by isolating the fractions having retention time about 26.2minutes (21.1 mg, 24.5% yield) which may have been isolated along withenantiomer and/or one or more diastereomers. The stereochemistry of thetitle compound was confirmed by ¹H-NMR nOe experiment. LC/MS (ES+APCI)m/z=398.1 (M+H).

Example 43

(4R,6R)-9-fluoro-4-hydroxy-13-oxa-2,16,20,21,24-pentaazapentacyclo[16.5.2.0^(2,6).0^(7,12).0^(21,25)]pentacosa-1(24),7,9,11,18(25),19,22-heptaen-17-one

The title compound was prepared according to the method of Example 41,substituting 2-chloroethylamine hydrochloride for(S)-1-amino-3-chloropropan-2-ol hydrochloride in Step A. The titlecompound was purified using a Chiral Tech OJ-H column, 20% EtOH inhexanes, by isolating fractions having a retention time of about 15.7minutes (10.7 mg, 14.2% yield). The stereochemistry of the titlecompound was confirmed by ¹H-NMR nOe experiment. LC/MS (ES+APCI)m/z=384.1 (M+H).

Example 43-B

(4R,6S)-9-fluoro-4-hydroxy-13-oxa-2,16,20,21,24-pentaazapentacyclo[16.5.2.0^(2,6).0^(7,12).0^(21,25)]pentacosa-1(24),7,9,11,18(25),19,22-heptaen-17-one

The title compound was isolated during the chiral separation reported inExample 43 by isolating fractions having a retention time of about 21.3minutes (15.9 mg, 21.1% yield) which may have been isolated along withthe enantiomer and/or one or more diastereomers. The stereochemistry ofthe title compound was confirmed by ¹H-NMR nOe experiment), LC/MS(ES+APCI) m/z=384.1 (M+H).

Example 44

4R,6R,15R)-9-fluoro-4,15-dihydroxy-13-oxa-2,17,21,22,25-pentaazapentacyclo[17.5.2.0^(2,6).0^(7,12).0^(22,26)]hexacosa-1(25),7(12),8,10,19(26),20,23-heptaen-18-one

The title compound was prepared according to the method of Example 41,substituting (R)-1-amino-3-chloropropan-2-ol hydrochloride (preparedaccording to the procedure described in Example 19, Step A using(R)-2-(chloromethyl)oxirane) for (S)-1-amino-3-chloropropan-2-olhydrochloride in Step A. The crude material was purified on a silica gelcolumn, eluting with CH₂Cl₂ to NH₄OH:MeOH:CH₂Cl₂ (0.5:5:95) (4 runs).Fractions containing the earlier eluting compound were collected toprovide 12 mg (10.9% yield) of the desired material. The stereochemistryof the title compound was confirmed by ¹H-NMR nOe experiment. LC/MS(ES+APCI) m/z=414.0 (M+H).

Example 44-B

(4R,6S,15R)-9-fluoro-4,15-dihydroxy-13-oxa-2,17,21,22,25-pentaazapentacyclo[17.5.2.0^(2,6).0^(7,12).0^(22,26)]hexacosa-1(25),7(12),8,10,19(26),20,23-heptaen-18-one

The title compound was isolated during the purification reported inExample 44. Fractions containing the later eluting compound werecollected to provide 15 mg (13.6% yield) of the title compound, whichmay have been isolated along with the enantiomer and/or one or morediastereomers. The stereochemistry of the title compound was confirmedby ¹H-NMR nOe experiment); LC/MS (ES+APCI) m/z=414.1 (M+H).

Example 45

Diastereomer 1 and Diastereomer 2 of(15S)-4,4,9-trifluoro-15-hydroxy-13-oxa-2,17,21,22,25-pentaazapentacyclo[17.5.2.0^(2,6).0^(7,12).0^(22,26)]hexacosa-1(25),7(12),8,10,19(26),20,23-heptaen-18-oneStep A: Preparation of (R)-5-(5-fluoro-2-methoxyphenyl)pyrrolidin-3-olhydrochloride

To solution of (R)-tert-butyl4-(tert-butyldimethylsilyloxy)-2-(5-fluoro-2-methoxyphenyl)pyrrolidine-1-carboxylate(1.01 g, 2.37 mmol) in CH₂Cl₂ (10 mL) at 0° C. was added 4 M HCl indioxane (5.93 mL, 23.7 mmol). The resulting mixture was warmed toambient temperature and stirred for 8 hours. The reaction mixture wasconcentrated under reduced pressure to give the crude material that wastriturated with ether. The resulting solids were filtered and driedunder reduced pressure to afford(R)-5-(5-fluoro-2-methoxyphenyl)pyrrolidin-3-ol hydrochloride (0.577 g,2.33 mmol, 98.2% yield). MS (APCI) m/z=212.0 (M+H).

Step B: Preparation of (R)-ethyl5-(2-(5-fluoro-2-methoxyphenyl)-4-hydroxypyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate

To a suspension of ethyl5-hydroxypyrazolo[1,5-a]pyrimidine-3-carboxylate (0.541 g, 2.61 mmol)and BOP reagent (1.57 g, 3.56 mmol) in DMF/CH₂Cl₂ (3 mL/3 mL) at 0° C.was added (R)-5-(5-fluoro-2-methoxyphenyl)pyrrolidin-3-ol hydrochloride(0.588 g, 2.37 mmol) followed by DIEA (1.66 mL, 9.50 mmol). Theresulting mixture was warmed to ambient temperature and stirred for 18hours. The reaction mixture was concentrated under reduced pressure togive the crude material that was diluted again with EtOAc (30 mL). Theorganic layer was washed with saturated aqueous NaHCO₃ followed bybrine, dried over MgSO₄, filtered, and concentrated under reducedpressure to give the crude material that was purified by silica gelflash column chromatography, eluting with CH₂Cl₂ to 5% MeOH in CH₂Cl₂ toafford (R)-ethyl5-(2-(5-fluoro-2-methoxyphenyl)-4-hydroxypyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate(0.735 g, 1.84 mmol, 77.3% yield). LC/MS (ES+APCI) m/z=401.1 (M+H).

Step C: Preparation of ethyl5-(2-(5-fluoro-2-methoxyphenyl)-4-oxopyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate

To a suspension of Dess-Martin periodinane (0.233 g, 0.549 mmol) inCH₂Cl₂ (2.2 mL) at 0° C. was added a solution of (R)-ethyl5-(2-(5-fluoro-2-methoxyphenyl)-4-hydroxypyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate(0.200 g, 0.499 mmol) in CH₂Cl₂ (1.5 mL). The resulting mixture waswarmed to ambient temperature and stirred for 18 hours. The reactionmixture was cooled to 0° C. and quenched with saturated aqueous NaHCO₃(5 mL) containing Na₂S₂O₃ (0.608 g, 3.85 mmol). The resulting mixturewas warmed to ambient temperature and stirred for 10 minutes. Theorganic layer was separated, washed with saturated aqueous NaHCO₃ (10mL) followed by brine (10 mL), dried over MgSO₄, filtered, andconcentrated under reduced pressure to afford ethyl5-(2-(5-fluoro-2-methoxyphenyl)-4-oxopyrrolidin-1-yl)pyrazolopyrimidine-3-carboxylate (0.164 g, 82.4% yield). LC/MS (ES+APCI)m/z=399.1 (M+H).

Step D: Preparation of ethyl5-(4,4-difluoro-2-(5-fluoro-2-methoxyphenyl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate

To a solution of ethyl5-(2-(5-fluoro-2-methoxyphenyl)-4-oxopyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate(0.162 g, 0.407 mmol) in CH₂Cl₂ (3 mL) was added a solution ofbis(2-methoxyethyl)aminosulfur trifluoride (0.134 mL, 0.691 mmol)followed by EtOH (0.00475 mL, 0.0813 mmol). The resulting mixture wasstirred at ambient temperature for 18 hours. The reaction mixture waspoured into saturated aqueous NaHCO₁ (6 mL) and extracted with CH₂Cl₂(2×10 mL). The combined organic layers were dried over MgSO₄, filtered,and concentrated under reduced pressure to give the crude material thatwas purified by silica column chromatography, eluting with 0-50%EtOAc/Hexanes to afford ethyl5-(4,4-difluoro-2-(5-fluoro-2-methoxyphenyl)pyrrolidin-1-yl)pyrazolopyrimidine-3-carboxylate (0.126 g, 65.6% yield). MS (APCI) m/z=420.9(M+H).

Step E: Preparation of5-(4,4-difluoro-2-(5-fluoro-2-hydroxyphenyl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylicacid

To a solution of ethyl5-(4,4-difluoro-2-(5-fluoro-2-methoxyphenyl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate(0.126 g, 0.267 mmol) in CH₂Cl₂ (1.3 mL) at 0° C. was added 1 M BBr₃ inCH₂Cl₂ (1.50 mL, 1.50 mmol). The resulting mixture was warmed to ambienttemperature and stirred for 18 hours. The reaction mixture was dilutedwith CH₂C17 (5 mL) and poured into a mixture of ice and saturatedaqueous NaHCO₃ (3 mL). The aqueous layer was then acidified to about pH3 with 1 N aqueous HCl. The aqueous layer was extracted with CH₂Cl₂(3×10 mL). The combined organic layers were dried over MgSO₄, filtered,and concentrated under reduced pressure to give a mixture of ethyl5-(4,4-difluoro-2-(5-fluoro-2-hydroxyphenyl)pyrrolidin-1-yl)pyrazolopyrimidine-3-carboxylate and5-(4,4-difluoro-2-(5-fluoro-2-hydroxyphenyl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylicacid. This mixture was taken up in MeOH-THE (0.25 mL/0.75 mL) at ambienttemperature, and 2 N aqueous LiOH (0.667 mL, 1.33 mmol) was added. Theresulting mixture was heated at 50° C. for 24 hours. The reactionmixture was cooled to ambient temperature and concentrated under reducedpressure to remove the organic solvents. The residue was diluted with 5mL of EtOAc and acidified to pH 3 to 4 with 6 N aqueous HCl withstirring. The organic layer was separated and the acidic aqueous layerwas extracted with EtOAc (2×5 mL). The combined organic layers weredried over MgSO₄, filtered, and concentrated under reduced pressure toafford5-(4,4-difluoro-2-(5-fluoro-2-hydroxyphenyl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylicacid (0.094 g, 93.1% yield). MS (APCI) m/z=378.9 (M+H).

Step F: Preparation ofN-((5)-3-chloro-2-hydroxypropyl)-5-(4,4-difluoro-2-(5-fluoro-2-hydroxyphenyl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

To a mixture of5-(4,4-difluoro-2-(5-fluoro-2-hydroxyphenyl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylicacid (0.047 g, 0.124 mmol) and HOBT (0.0252 g, 0.186 mmol) in DMF (1 mL)at ambient temperature was added EDCI (0.0357 g, 0.186 mmol). Theresulting mixture was stirred for 1 hour. To this mixture was added(5)-1-amino-3-chloropropan-2-ol hydrochloride (Example 19, Step A;0.0218 g, 0.149 mmol) followed by DIEA (0.0656 mL, 0.373 mmol) atambient temperature. The resulting mixture was stirred for 48 hours. Thereaction mixture was diluted with EtOAc (10 mL), and the organic layerwas washed with a 1:1 mixture of brine and water. The aqueous layer wasseparated and extracted with EtOAc (2×10 mL). The combined organiclayers were washed with a 1:1 mixture of brine and water (15 mL) andcombined with the organic layer obtained previously. The organic layerwas dried over MgSO₄, filtered, and concentrated under reduced pressureto affordNAS)-3-chloro-2-hydroxypropyl)-5-(4,4-difluoro-2-(5-fluoro-2-hydroxyphenyl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide(0.061 g, 105% yield). LC/MS (ES+APCI) m/z=468.1 (M−H).

Step G: Preparation of Diastereomers 1 and 2 of(15S)-4,4,9-trifluoro-15-hydroxy-13-oxa-2,17,21,22,25-pentaazapentacyclo[17.5.2.0^(2,6).0^(7,12).0^(22,26)]hexacosa-1(25),7(12), 8,10,19(26),20,23-heptaen-18-one

A mixture ofN—((S)-3-chloro-2-hydroxypropyl)-5-(4,4-difluoro-2-(5-fluoro-2-hydroxyphenyl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide(0.060 g, 0.128 mmol) and Cs₂CO₃ (0.208 g, 0.639 mmol) in DMF (6.4 mL)was heated at 85° C. for 30 minutes. The reaction mixture was cooled toambient temperature and filtered. The filtrate was concentrated underreduced pressure to give the crude material which was purified by silicagel flash column chromatography (CH₂Cl₂ to NH₄OH:MeOH:CH₂C19=0.5:5:95)to afford a mixture of the diastereomers. The isolated diastereomerswere further purified by chiral column chromatography (Chiral Tech OD-Hcolumn, 20% EtOH in hexanes). Fractions having a retention time of about17.1 minutes were isolated to afford the title compound designated asDiastereomer 1 (11 mg, 20% yield.; MS (APCI) m/z=434.2 (M+H). Fractionshaving a retention time of about 21.0 minutes were isolated to providethe title compound designated as Diastereomer 2 (13 mg; 24% yield); MS(APCI) m/z=434.2 (M+H).

1. (canceled)
 2. A pharmaceutical composition comprising a compoundselected from the group consisting of:(6R)-9-fluoro-15-hydroxy-13-oxa-2,11,17,21,22,25-hexaazapentacyclo[17.5.2.0^(2,6).0^(7,12).0^(22,26)]hexacosa-1(25),7,9,11,19(26),20,23-heptaen-18-one(6R,15R)-9-fluoro-15-hydroxy-13-oxa-2,11,17,21,22,25-hexaazapentacyclo-[17.5.2.0^(2,6).0^(7,12).0^(22,26)]hexacosa-1(25),7,9,11,19(26),20,23-heptaen-18-one;(6R)-9-fluoro-13-oxa-2,11,16,20,21,24-hexaazapentacyclo[16.5.2.0^(2,6).0^(7,12).0^(21,25)]pentacosa-1(24),7,9,11,18(25),19,22-heptaen-17-one;(6R)-9-fluoro-13-oxa-2,11,18,22,23,26-hexaazapentacyclo[18.5.2.0^(2,6).0^(7,12).0^(23,27)]heptacosa-1(26),7,9,11,20(27),21,24-heptaen-19-one;(6R)-9-fluoro-2,11,13,16,20,21,24-heptaazapentacyclo[16.5.2.0^(2,6).0^(7,12).0^(21,25)]pentacosa-1(24),7,9,11,18(25),19,22-heptaen-17-one;(6R)-9-fluoro-2,11,13,17,21,22,25-heptaazapentacyclo[17.5.2.0^(2,6).0^(7,12).0^(22,26)]hexacosa-1(25),7,9,11,19(26),20,23-heptaen-18-one;(6R)-9-fluoro-17-methyl-13-oxa-2,11,17,21,22,25-hexaazapentacyclo[17.5.2.0^(2,6).0^(7,12).0^(22,26)]hexacosa-1(25),7,9,11,19(26),20,23-heptaen-18-one;(6R)-9,15,15-trifluoro-13-oxa-2,11,17,21,22,25-hexaazapentacyclo[17.5.2.0^(2,6).0^(7,12).0^(22,26)]hexacosa-1(25),7,9,11,19(26),20,23-heptaen-18-one;(6R)-9-fluoro-2,11,16,20,21,24-hexaazapentacyclo[16.5.2.0^(2,6).0^(7,12).0^(21,25)]pentacosa-1(24),7,9,11,18(25),19,22-heptaen-17-one;(6R)-9-fluoro-15-methyl-2,11,16,20,21,24-hexaazapentacyclo[16.5.2.0^(2,6).0^(7,12).0^(21,25)]pentacosa-1(24),7,9,11,18(25),19,22-heptaen-17-one;(6R)-9-fluoro-15,15-dimethyl-13-oxa-2,11,17,21,22,25-hexaazapentacyclo[17.5.2.0^(2,6).0^(7,12).0^(22,26)]hexacosa-1(25),7,9,11,19(26),20,23-heptaen-18-one;and(6R)-9-fluoro-15,15-dimethyl-2,11,16,20,21,24-hexaazapentacyclo[16.5.2.0^(2,6).0^(7,12).0^(21,25)]pentacosa-1(24),7,9,11,18(25),19,22-heptaen-17-one;or a pharmaceutically acceptable salt thereof and a pharmaceuticallyacceptable diluent or carrier.
 3. The pharmaceutical composition ofclaim 2, wherein the compound is(6R)-9-fluoro-15-hydroxy-13-oxa-2,11,17,21,22,25-hexaazapentacyclo[17.5.2.0^(2,6).0^(7,12).0^(22,26)]hexacosa-1(25),7,9,11,19(26),20,23-heptaen-18-one,or a pharmaceutically acceptable salt thereof.
 4. The pharmaceuticalcomposition of claim 2, wherein the compound is(6R,15R)-9-fluoro-15-hydroxy-13-oxa-2,11,17,21,22,25-hexaazapentacyclo-[17.5.2.0^(2,6).0^(7,12).0^(22,26)]hexacosa-1(25),7,9,11,19(26),20,23-heptaen-18-one,or a pharmaceutically acceptable salt thereof.
 5. The pharmaceuticalcomposition of claim 2, wherein the compound is(6R)-9-fluoro-13-oxa-2,11,16,20,21,24-hexaazapentacyclo[16.5.2.0^(2,6).0^(7,12).0^(21,25)]pentacosa-1(24),7, 9,11,18(25),19,22-heptaen-17-one,or a pharmaceutically acceptable salt thereof.
 6. The pharmaceuticalcomposition of claim 2, wherein the compound is(6R)-9-fluoro-13-oxa-2,11,18,22,23,26-hexaazapentacyclo[18.5.2.0^(2,6).0^(7,12).0^(23,27)]heptacosa-1(26),7,9,11,20(27),21,24-heptaen-19-one,or a pharmaceutically acceptable salt thereof.
 7. The pharmaceuticalcomposition of claim 2, wherein the compound is(6R)-9-fluoro-2,11,13,16,20,21,24-heptaazapentacyclo[16.5.2.0^(2,6).0^(7,12).0^(21,25)]pentacosa-1(24),7,9,11,18(25),19,22-heptaen-17-one,or a pharmaceutically acceptable salt thereof.
 8. The pharmaceuticalcomposition of claim 2, wherein the compound is(6R)-9-fluoro-2,11,13,17,21,22,25-heptaazapentacyclo[17.5.2.0^(2,6).0^(7,12).0^(22,26)]hexacosa-1(25),7,9,11,19(26),20,23-heptaen-18-one,or a pharmaceutically acceptable salt thereof.
 9. The pharmaceuticalcomposition of claim 2, wherein the compound is(6R)-9-fluoro-17-methyl-13-oxa-2,11,17,21,22,25-hexaazapentacyclo[17.5.2.0^(2,6).0^(7,12).0^(22,26)]hexacosa-1(25),7,9,11,19(26),20,23-heptaen-18-one,or a pharmaceutically acceptable salt thereof.
 10. The pharmaceuticalcomposition of claim 2, wherein the compound is(6R)-9,15,15-trifluoro-13-oxa-2,11,17,21,22,25-hexaazapentacyclo[17.5.2.0^(2,6).0^(7,12).0^(22,26)]hexacosa-1(25),7,9,11,19(26),20,23-heptaen-18-one,or a pharmaceutically acceptable salt thereof.
 11. The pharmaceuticalcomposition of claim 2, wherein the compound is(6R)-9-fluoro-2,11,16,20,21,24-hexaazapentacyclo[16.5.2.0^(2,6).0^(7,12).0^(21,25)]pentacosa-1(24),7,9,11,18(25),19,22-heptaen-17-one,or a pharmaceutically acceptable salt thereof.
 12. The pharmaceuticalcomposition of claim 2, wherein the compound is(6R)-9-fluoro-15-methyl-2,11,16,20,21,24-hexaazapentacyclo[16.5.2.0^(2,6).0^(7,12).0^(21,25)]pentacosa-1(24),7,9,11,18(25),19,22-heptaen-17-one,or a pharmaceutically acceptable salt thereof.
 13. The pharmaceuticalcomposition of claim 2, wherein the compound is(6R)-9-fluoro-15,15-dimethyl-13-oxa-2,11,17,21,22,25-hexaazapentacyclo[17.5.2.0^(2,6).0^(7,12).0^(22,26)]hexacosa-1(25),7,9,11,19(26),20,23-heptaen-18-one,or a pharmaceutically acceptable salt thereof.
 14. The pharmaceuticalcomposition of claim 2, wherein the compound is(6R)-9-fluoro-15,15-dimethyl-2,11,16,20,21,24-hexaazapentacyclo[16.5.2.0^(2,6).0^(7,12).0^(21,25)]pentacosa-1(24),7,9,11,18(25),19,22-heptaen-17-one,or a pharmaceutically acceptable salt thereof.
 15. The pharmaceuticalcomposition of claim 2, wherein the compound is selected from the groupconsisting of:(6R)-9-fluoro-15-hydroxy-13-oxa-2,11,17,21,22,25-hexaazapentacyclo[17.5.2.0^(2,6).0^(7,12).0^(22,26)]hexacosa-1(25),7,9,11,19(26),20,23-heptaen-18-one;(6R,15R)-9-fluoro-15-hydroxy-13-oxa-2,11,17,21,22,25-hexaazapentacyclo-[17.5.2.0^(2,6).0^(7,12).0^(22,26)]hexacosa-1(25),7,9,11,19(26),20,23-heptaen-18-one;(6R)-9-fluoro-13-oxa-2,11,16,20,21,24-hexaazapentacyclo[16.5.2.0^(2,6).0^(7,12).0^(21,25)]pentacosa-1(24),7,9,11,18(25),19,22-heptaen-17-one;(6R)-9-fluoro-13-oxa-2,11,18,22,23,26-hexaazapentacyclo[18.5.2.0^(2,6).0^(7,12).0^(23,27)]heptacosa-1(26),7,9,11,20(27),21,24-heptaen-19-one;(6R)-9-fluoro-2,11,13,16,20,21,24-heptaazapentacyclo[16.5.2.0^(2,6).0^(7,12).0^(21,25)]pentacosa-1(24),7,9,11,18(25),19,22-heptaen-17-one;(6R)-9-fluoro-2,11,13,17,21,22,25-heptaazapentacyclo[17.5.2.0^(2,6).0^(7,12).0^(22,26)]hexacosa-1(25),7,9,11,19(26),20,23-heptaen-18-one;(6R)-9-fluoro-17-methyl-13-oxa-2,11,17,21,22,25-hexaazapentacyclo[17.5.2.0^(2,6).0^(7,12).0^(22,26)]hexacosa-1(25),7,9,11,19(26),20,23-heptaen-18-one;(6R)-9,15,15-trifluoro-13-oxa-2,11,17,21,22,25-hexaazapentacyclo[17.5.2.0^(2,6).0^(7,12).0^(22,26)]hexacosa-1(25),7,9,11,19(26),20,23-heptaen-18-one;(6R)-9-fluoro-2,11,16,20,21,24-hexaazapentacyclo[16.5.2.0^(2,6).0^(7,12).0^(21,25)]pentacosa-1(24),7,9,11,18(25),19,22-heptaen-17-one;(6R)-9-fluoro-15-methyl-2,11,16,20,21,24-hexaazapentacyclo[16.5.2.0^(2,6).0^(7,12).0^(21,25)]pentacosa-1(24),7,9,11,18(25),19,22-heptaen-17-one;(6R)-9-fluoro-15,15-dimethyl-13-oxa-2,11,17,21,22,25-hexaazapentacyclo[17.5.2.0^(2,6).0^(7,12).0^(22,26)]hexacosa-1(25),7,9,11,19(26),20,23-heptaen-18-one;and(6R)-9-fluoro-15,15-dimethyl-2,11,16,20,21,24-hexaazapentacyclo[16.5.2.0^(2,6).0^(7,12).0^(21,25)]pentacosa-1(24),7,9,11,18(25),19,22-heptaen-17-one.16. The composition of claim 2, wherein the compound is in the form of apharmaceutically acceptable salt.